KR20210091705A - Human Amylin Analog Polypeptides and Methods of Use - Google Patents

Abstract

The present invention relates to isolated polypeptides that are analogs of human amylin. The disclosed amylin analog polypeptides have advantageous physicochemical properties compared to endogenous amylin, such as a longer elimination half-life (t _1/2 ) and improved solubility and thermal stability. The invention also relates to methods of using the disclosed amylin analog polypeptides of the invention as well as methods of producing them in various therapeutic indications. The disclosed amylin analog polypeptides are particularly useful in methods of treating metabolic diseases or disorders, such as type 1 and type 2 diabetes, and providing weight loss.

Description

Human Amylin Analog Polypeptides and Methods of Use

This application claims priority and benefit to U.S. Provisional Application No. 62/744,236, filed on October 11, 2018, which is incorporated herein by reference in its entirety.

sequence list

This application is filed electronically in ASCII format and includes a Sequence Listing, which is incorporated herein by reference in its entirety. The ASCII copy, created on October 10, 2019, is named 616782_102487-055PC_SL-10-10-2019.txt and has a capacity of 123,647 bytes.

technical field

The present invention relates to isolated polypeptides that are analogs of human amylin. The disclosed amylin analog polypeptides have advantageous physicochemical properties compared to endogenous amylin, such as a longer elimination half-life (t _1/2 ) and improved solubility and thermal stability. The invention also relates to methods of using the disclosed amylin analog polypeptides of the invention as well as methods of producing them in various therapeutic indications. As described in more detail below, the disclosed amylin analog polypeptides are particularly useful in methods of treating metabolic diseases or disorders, such as type 1 and type 2 diabetes, and providing weight loss.

Human amylin, or islet amyloid polypeptide (IAPP), is a 37-residue polypeptide hormone. Amylin is secreted along with insulin from pancreatic β-cells in a ratio of approximately 100:1 (insulin:amylin). Pro-islet amyloid polypeptide (i.e., pro-IAPP) is a 67 amino acid pro-peptide of 7404 Daltons produced in pancreatic β-cells that undergoes post-translational modifications involving protease cleavage. Residual amylin is produced. Loss of β-cell function, which occurs early in type 1 diabetes and late in type 2 diabetes, results in a deficiency of insulin and amylin secretion.

Amylin acts as part of the endocrine pancreas, the corresponding cells within the pancreas that synthesize and secrete hormones. Amylin contributes to glycemic control; It is secreted from the pancreatic islets into the blood circulation and is cleared by peptidase in the kidneys. The metabolic function of amylin is well characterized as an inhibitor of the appearance of nutrients such as glucose in plasma. It thus acts as a synergist to insulin, a peptide that regulates blood sugar levels and regulates the distribution and absorption of glucose in the body. The role of insulin in the body is, among other things, to prevent blood sugar levels from rising too high, especially after a meal.

Amylin is believed to play a role in regulating glycemia by slowing gastric emptying and promoting a feeling of fullness (ie, feeling full), thereby preventing peak postprandial (ie, post-meal) blood sugar levels. The overall effect is to slow the rate of glucose appearance in the blood after feeding. Amylin also lowers the secretion of glucagon by the pancreas. Glucagon's role in the body is, among other things, to prevent blood sugar levels from dropping too low. This is significant because certain type 1 diabetes mellitus, for example, tend to secrete excess blood sugar-elevating glucagon only after a meal.

For a number of reasons, human amylin, with a serum half-life of about 13 minutes, cannot be used as a therapeutic agent. Rather, pramlintide (Symlin®, developed by Amylin Pharmaceuticals, Inc. (San Diego, CA) and marketed by AstraZeneca plc (Cambridge, UK)) is used despite optimal insulin therapy. , developed as a synthetic analogue of human amylin for the treatment of patients with type 1 or type 2 diabetes who use dietary insulin but cannot achieve desired glycemic control. Pramlintide differs from human amylin in 3 of its 37 amino acids. These modifications provide pramlintide with a longer half-life of approximately 48 minutes in humans and reduce its tendency to aggregation characteristically found in human amylin.

For the treatment of type 1 diabetes, pramlintide is administered up to 4 times per day by subcutaneous injection before meals as an adjunct to postprandial insulin therapy. Pramlintide is immiscible with insulin; A separate syringe is used. Reported side effects of pramlintide include nausea and vomiting. Adverse reactions may include severe hypoglycemia, particularly type 1 diabetes. Consequently, the dosage of prandial insulin is reduced for patients initiating administration of pramlintide.

For the treatment of type 2 diabetes mellitus, pramlintide is administered by subcutaneous injection before each meal at the recommended starting dose, which is gradually increased to the target maintenance dose. Another under study analog of human amylin, davalintide (AC2307; also developed by Amylin Pharmaceuticals, Inc.), has a half-life of about 26 minutes.

Accordingly, a need exists for improved amylin analog polypeptides in which mimetic amylin activity has greater therapeutic potential than endogenous human amylin and existing amylin analogs, such as both pramlintide and davalintide. do.

It has now been discovered that the polypeptides of the present invention, and pharmaceutically acceptable compositions thereof, are effective as amylin analogs. Such a polypeptide has SEQ ID NO:199, or a pharmaceutically acceptable salt thereof, of the general formula:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TX ₃₇ -(OH/NH ₂ )(SEQ ID NO: 199), where:

X ₁ is S, K, k, H or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; X ₈ is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₃ is A, S, E or K; X ₁₄ is N, n, d, Y or Q; X ₁₅ is E, F, f, Y, I, k, K or α-aminoisobutyric acid (Aib); X ₁₆ is k, K, L, Aib, N -methyl leucine ( N -MeL) or 1; X ₁₇ is H, V, Q, R, k, K or Aib; X ₁₈ is K, H or R; X ₁₉ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ is P, R or K; X ₃₁ is k, K, N or H; X ₃₅ is e, E, N, K, G, A, Y or P; and X ₃₇ is Y or P;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge;

provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K.

1 shows specific reference polypeptides: human amylin, SEQ ID NO: 300; rat amylin, SEQ ID NO: 301; pramlintide, SEQ ID NO:302; Davalintide, SEQ ID NO:303; hCT (human calcitonin), SEQ ID NO: 304; sCT (salmon calcitonin), SEQ ID NO: 305; and a table showing a comparative sequence alignment to beta-calcitonin-gene-related peptide (β-CGRP), SEQ ID NO:306.
2A, 2B, 2C and 2D show amylin analogue A73 acylated at hAMY3R (Fig. 2A), amylin analogue A73 acylated at hCTR (Fig. 2B), hAMY3R pramlintide (Fig. 2C), and A diagram depicting a dose-response curve for human calcitonin (hCalcitonin) in hCTR ( FIG. 2D ). The term “acylated” as used herein, with respect to a disclosed polypeptide, means that the disclosed polypeptide is each optionally substituted with one or more lipophilic substituents via a spacer, provided that “lipophilic substituents” and “ A "spacer" is defined herein.
3A and 3B show polypeptide clearance from the kidney after intravenous infusion of a linear, ie, non-acylated, polypeptide ( FIG. 3A ) and a conjugated, ie, acylated, polypeptide ( FIG. 3B ). Data from pharmacokinetic studies to evaluate (CL).

1. General Description of Certain Embodiments of the Invention

The present invention relates to isolated polypeptides that are amylin analogs as well as pharmaceutical compositions comprising these polypeptides. The invention also relates to methods of making and using such amylin analog polypeptides. These amylin analog polypeptides are particularly useful in methods of treating metabolic diseases or disorders, such as type 1 and type 2 diabetes, obesity, and methods of providing weight loss.

2. Definition

It should be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. As used in this specification and the appended claims, the singular forms include the plural referents unless clearly indicated otherwise. Thus, for example, reference to "a solvent" includes a combination of two or more such solvents, reference to "peptide" includes one or more peptides, or mixtures of peptides, and reference to "drug" Reference includes one or more drugs, reference to “osmotic delivery device” includes one or more osmotic delivery devices, and the like. Unless specifically stated or clear from the context, the term “or” as used herein is to be understood inclusive and encompasses both “or” and “and”.

Unless specifically stated or clear from context, the term “about” as used herein is understood to be within the normal tolerance of the art, eg, within two standard deviations of the mean. about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05% or 0.01% of the stated value. can be understood Unless otherwise clear from context, all numerical values provided herein are modified by the term “about.”

Unless specifically stated or clear from the context, the term “substantially” as used herein is understood to be within the narrow range of variation in the art or otherwise normal tolerances. substantially within 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, 0.01% or 0.001% of the stated value.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although other methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred materials and methods are described herein.

In describing and claiming the present invention, the following terms will be used in accordance with the definitions set forth below.

The terms “drug”, “therapeutic agent” and “beneficial agent” are used interchangeably to refer to any therapeutically active substance that is delivered to a subject to produce a desired beneficial effect. In one embodiment of the invention, the drug is a polypeptide. In another embodiment of the invention, the drug is a small molecule, eg, a hormone, eg, an androgen or an estrogen. The devices and methods of the present invention are well suited for the delivery of proteins, small molecules and combinations thereof.

The terms “peptide,” “polypeptide,” and “protein” are used interchangeably herein, and typically include two or more amino acids (eg, most typically L-amino acids, also, eg, D- amino acids, modified amino acids, amino acid analogs, and amino acid mimetics).

In some embodiments, naturally occurring L-amino acids are represented by the conventional three-letter, or uppercase, one-letter amino acid designation in Table 1. In another embodiment, the naturally occurring L-amino acids and D-amino acids are both represented by the conventional three-letter, or uppercase, one-letter notation in Table 1. In another embodiment, the D-amino acid is a lowercase one-letter amino acid notation corresponding to the one-letter notation in Table 1, i.e. g, a, l, m, f, w, k, q, e, s, p, v , i, c, y, h, r, n, d and t.

Peptides may be of natural origin, synthetically produced, or recombinantly expressed. Peptides may also contain additional groups that modify the amino acid chain, for example, functional groups added via post-translational modifications. Examples of post-translational modifications include acetylation, alkylation (including methylation), biotinylation, glutamylation, glycylation, glycosylation, isoprenylation, lipoylation, phosphopantetheinylation, phosphorylation, selenization and C-terminus. including, but not limited to, amidation of The term peptide also includes peptides comprising modifications of the amino terminus and/or the carboxy terminus. Modifications of the terminal amino group include, but are not limited to, des-amino, N-lower alkyl, N-di-lower alkyl and N-acyl modifications. Modifications of terminal carboxy groups include, but are not limited to, amide, lower alkyl amide, dialkyl amide and lower alkyl ester modifications (eg, lower alkyl is C1-C4 alkyl). The term peptide also includes modifications as described above, but not limited to, of amino acids that fall between the amino terminus and the carboxy terminus. In one embodiment, the peptide may be modified by the addition of a small molecule drug.

The terminal amino acid at one end of the peptide chain typically has a free amino group (ie, the amino terminus). The terminal amino acid at the other end of the chain typically has a free carboxyl group (ie, the carboxy terminus). Typically, the amino acids making up the peptide are numbered in increasing order starting at the amino terminus and moving towards the carboxy terminus of the peptide.

The phrase “amino acid residue” as used herein refers to an amino acid incorporated into a peptide by an amide bond or an amide bond mimetic.

As used herein, the term "insulinotropic" typically refers to a compound, e.g., a peptide, that stimulates or affects the production and/or activity of insulin (e.g., insulinotropic hormone). refers to ability. Such compounds typically stimulate or otherwise affect the secretion or biosynthesis of insulin in a subject. Thus, an “insulin affinity peptide” is an amino acid-containing molecule capable of stimulating or otherwise affecting the secretion or biosynthesis of insulin.

The term "insulin affinity peptide" as used herein refers to glucagon-like peptide 1 (GLP-1) as well as derivatives and analogs thereof, GLP-1 receptor agonists such as exenatide, the amino acid sequence of SEQ ID NO: 307 Exenatide with, as well as derivatives and analogs thereof, but are not limited thereto.

The term "acylated" as used herein, with respect to a disclosed polypeptide, means that the disclosed polypeptide is each substituted with one or more lipophilic substituents, optionally via a spacer, provided that "lipophilic substituents" and " A "spacer" is defined herein. Certain lipophilic substituents are each capable of binding to albumin through a spacer and conferring affinity to the albumin for the resulting acylated polypeptide. The extent is variable and depends on a number of factors that each lipophilic substituent binds to albumin through a spacer and confers affinity to the albumin for the resulting acylated polypeptide. Numerous factors include the identity of lipophilic substituents, optional spacers, and sites of covalent attachment to polypeptides and polypeptides.

The term "linear" or "linear polypeptide" as used herein refers to a "non-acylated" polypeptide, i.e., the disclosed amylin analogs each optionally lacking a lipophilic substituent via a spacer, “Lipophilic substituents” and “spacers” are defined herein.

The term "conjugated" or "conjugated polypeptide" as used herein refers to an "acylated" polypeptide, i.e., the disclosed amylin analogs each optionally having one or more lipophilic substituents via a spacer. provided that “lipophilic substituents” and “spacers” are defined herein.

As used herein, the term "pharmaceutically acceptable salt" is suitable for use in contact with tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc. within the scope of sound medical judgment, and is reasonably harmful. / refers to the corresponding salt proportional to the benefit ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., J. Pharmaceutical Sciences, 1977, 66, 1-19 describe the pharmaceutically acceptable salts detailed above. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, non-toxic acid addition salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or organic acids such as acetic acid, trifluoroacetic acid (TFA), oxalic acid, maleic acid, salts of amino groups formed with tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate , digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy -ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate , palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate , undecanoate, valerate salts and the like.

Salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium and N+(C1-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Additional pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates. amine cations formed using

The phrase “incretin mimic” as used herein includes: a GLP-1 peptide, a GLP-1 receptor agonist, a peptide derivative of GLP-1, a peptide analog of GLP-1; exenatide, exenatide having the amino acid sequence of SEQ ID NO: 307, exenatide peptides, peptide derivatives of exenatide, and peptide analogs of exenatide, but are not limited thereto. Examples of preferred incretin mimetics include exenatide, exenatide having the amino acid sequence of exendin-4 (a naturally occurring form of exenatide), exenatide-LAR, lixisenatide, GLP-1 (7 -36), liraglutide, semaglutide, dulaglutide, albiglutide and taspoglutide. Incretin mimetics are also referred to herein as "insulin affinity peptides". Incretin mimetics that target the GLP-1 receptor are also known in the literature as "GLP-1 receptor agonists" or "GLP-1 agonists", the two terms being used interchangeably herein.

The term "exenatide" as used herein refers to exenatide, exenatide having the amino acid sequence of SEQ ID NO: 307 of _{(HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-NH 2 ), native exendin-4, exenatide peptide, exena} Tide peptide analogs and exenatide peptide derivatives.

The term “GLP-1” refers to a polypeptide produced by L-cells that is located primarily in the ileum and colon and to a lesser extent by L-cells in the duodenum and jejunum. GLP-1 is a regulatory peptide that binds to the extracellular domain of the G-binding protein receptor, the GLP-1 receptor (GLP-1R), on β cells and via adenyl cyclase activity, and the production of cAMP is Stimulates the insulin response to nutrients [Baggio 2007, "Biology of incretins: GLP-1 and GIP," Gastroenterology, vol. 132(6):2131-57; Holst 2008, "The incretin system and its role in 2 diabetes mellitus," Mol Cell Endocrinology, vol. 297(1-2):127-36]. The effects of GLP-1R agonism vary. GLP-1 maintains glucose homeostasis by enhancing endogenous glucose-dependent insulin secretion, provides β-cell glucose that is competent and sensitive to GLP-1, inhibits glucagon release, restores phase 1 and phase 2 insulin secretion, It exhibits gastric emptying, reduces food intake, and increases satiety [Holst 2008 Mol. Cell Endocrinology; Kjems 2003 "The influence of GLP-1 on glucose-stimulated insulin secretion: effects on beta-cell sensitivity in type 2 and nondiabetic subjects," Diabetes, vol. 52(2): 380-86; Holst 2013 “Incretin hormones and the satiation signal,” Int J Obes (Lond), vol. 37(9):1161-69; Seufert 2014, "The extra-pancreatic effects of GLP-1 receptor agonists: a focus on the cardiovascular, gastrointestinal and central nervous systems," Diabetes Obes Metab, vol. 16(8): 673-88]. The risk of hypoglycemia is minimal given the mode of action of GLP-1.

As described in more detail below, in some embodiments, an amylin analog polypeptide disclosed herein is provided in a method of treating type 1 diabetes as an adjunct to treatment with insulin. The term “insulin” as used herein refers to human insulin or any insulin analogue. Exemplary non-limiting insulin analogs include those listed in Table 2:

The term "meal insulin" as used herein refers to a fast-acting insulin formulation that reaches peak blood concentrations at approximately 45 to 90 minutes and peak activity approximately 1 to 3 hours after administration, at or near mealtime. is administered to

"Vehicle" as used herein refers to a medium used to deliver a compound, eg, a drug, or particles containing a drug. Vehicles of the present invention typically include polymers, such as polymers and solvents. Suspension vehicles of the present invention typically comprise the solvent and polymer used to prepare suspension formulations further comprising drug particle formulations.

The phrase “phase separation” as used herein refers to the formation of multiple phases (eg, liquid and gel phases) in a suspension vehicle, eg, when the suspension vehicle is contacted with an aqueous environment. In some embodiments of the invention, the suspension vehicle is formulated to exhibit phase separation upon contact with an aqueous environment having less than approximately 10% water.

The phrase “single-phase” as used herein refers to a solid, semi-solid or liquid homogeneous system that is physically and chemically uniform throughout.

The term “dispersed” as used herein refers to dissolving, dispersing, suspending or otherwise distributing a compound, eg, a drug particle formulation, in a suspension vehicle.

The phrase “chemically stable,” as used herein, refers to the formation in a formulation of an acceptable percentage of degradation products over a defined period of time by chemical pathways such as deamidation (usually by hydrolysis), aggregation or oxidation. refers to

The phrase “physically stable,” as used herein, refers to the formation in the formulation of an acceptable percentage of aggregates (eg, dimers and other higher molecular weight products). In addition, a physically stable formulation does not change its physical state, for example, from liquid to solid, or from amorphous to crystalline form.

The term “viscosity” as used herein typically refers to a value determined from the ratio of shear stress to shear rate essentially as follows (e.g., Considine, DM & Considine, GD, Encyclopedia of Chemistry) , 4th Edition, Van Nostrand, Reinhold, NY, 1984]:

where F/A = shear stress (force per unit area),

μ = constant of proportionality (viscosity), and

V/L = rate per layer thickness (shear rate).

From this relationship, the ratio of shear stress to shear rate determines the viscosity. Measurements of shear stress and shear rate are typically determined using parallel plate rheometry performed under selected conditions (eg, a temperature of about 37° C.). Another method for the determination of viscosity is the measurement of kinematic viscosity using a viscometer, such as a Cannon-Fenske viscometer, a Ubbelohde viscometer for Cannon-Fenske opaque solutions, or an Ostwald viscometer. In general, the suspension vehicles of the present invention have sufficient viscosity to prevent settling of the particle formulation suspended therein during storage and for use in delivery methods, for example, in implantable drug delivery devices.

As used herein, the term “non-aqueous” means, for example, about 10% by weight or less, such as about 7% by weight or less, about 5% by weight or less, and/or refers to a total moisture content of about 4% by weight or less. In addition, the particle formulations of the present invention contain no more than about 10% by weight residual moisture, such as no more than about 5% by weight.

The term “subject” as used herein includes humans and non-human primates such as rhesus macaques and other primates, including other monkey species and chimpanzees and other ape species; farm animals such as cattle, sheep, pigs, goats and horses; domestic mammals such as dogs and cats; laboratory animals including rodents such as mice, rats and guinea pigs; refers to any member of the subfamily Chordoptera, including, but not limited to, birds, including domestic, wild and wild birds, such as chickens, turkeys and other poultry, ducks, geese, and the like. The term does not denote a specific age or gender. Thus, it is intended to encompass both adult and neonatal individuals.

As used herein, the terms “treatment”, “treat” and “treating” refer to a disease or disorder as described herein, or one or more symptoms thereof, which reverses, alleviates, or delays the onset of refers to In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment can be administered to a subject predisposed to the onset of symptoms (eg, in light of a history of symptoms and/or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.

The term “osmotic delivery device” as used herein refers to a device typically used for delivery of a drug (eg, a disclosed amylin analog polypeptide) to a subject, wherein the device includes, for example, a drug ( For example, a reservoir (e.g., made of a titanium alloy) having a lumen containing a suspension formulation comprising the disclosed amylin analog polypeptide) and an osmotic agent formulation. A piston assembly positioned in the lumen isolates the suspension formulation from the osmotic formulation. The semipermeable membrane is positioned at a first distal end of the reservoir adjacent the osmotic agent formulation, and a diffusion regulator (defining a delivery orifice through which the suspension formulation exits the device) is positioned at the second distal end of the reservoir adjacent the suspension formulation. Typically, an osmotic delivery device is implanted in a subject, eg, subdermally or subcutaneously (eg, in the interior, exterior or posterior and abdominal regions of the upper arm). An exemplary osmotic delivery device is a DUROS® (ALZA Corporation, Mountain View, CA) delivery device. Examples of terms synonymous with "osmotic delivery device" include "osmotic drug delivery device", "osmotic drug delivery system", "osmotic device", "osmotic delivery device", "osmotic delivery system", "osmotic pump", "implantation" possible drug delivery devices", "drug delivery systems", "drug delivery devices", "implantable osmotic pumps", "implantable drug delivery systems", and "implantable delivery systems". Other terms for "osmotic delivery device" are known in the art.

The term "continuous delivery" as used herein refers to substantially continuous release of a drug, typically from an osmotic delivery device and into tissues near the site of implantation, such as subdermal and subcutaneous tissue. For example, an osmotic delivery device releases a drug at an essentially predetermined rate based on the principle of osmotic pressure. The extracellular fluid enters the osmotic delivery device directly into the osmotic engine through the semipermeable membrane, causing the piston to expand to drive it at a slow and consistent rate of movement. The movement of the piston forces the drug formulation to be released through the orifice of the diffusion regulator. Thus, drug release from the osmotic delivery device is slow, controlled, and continuous at a rate.

The term "substantially steady state delivery" as used herein typically refers to drug delivery at or near a target concentration over a defined period of time, wherein the amount of drug being delivered from an osmotic delivery device is substantially zero order delivery. . Substantial zero-order delivery of an active agent (eg, a disclosed amylin analog polypeptide) results in a constant drug rate delivered and independent of the drug available in the delivery system; For example, for zero-order delivery, if the rate of drug delivered is graphed versus time and a line is fitted to the data, the line will have a slope of approximately zero as determined by standard methods (eg, linear regression). means to have

The phrase “drug half-life” as used herein refers to how long it takes for a drug to be cleared from plasma to half its concentration. The half-life of a drug is usually measured by monitoring how the drug degrades when administered via injection or intravenously. Drugs are usually detected using, for example, radioimmunoassay (RIA), chromatographic methods, electrochemical luminescence (ECL) assays, enzyme linked immunosorbent assays (ELISA) or immunoenzyme sandwich assays (IEMA).

The terms “μg” and “mcg” and “ug” are understood to mean “microgram”. Similarly, the terms “μL” and “uL” are understood to mean “microliter”, and the terms “μM” and “uM” are understood to mean “micromolar”.

The term “serum” means any blood product in which a substance can be detected. Accordingly, the term serum includes at least whole blood, serum and plasma. For example, "amount of [substance] in a subject's serum" encompasses "amount of [substance] in plasma of a subject."

Criteria is defined as the last evaluation on or before the date of initial placement of the osmotic delivery device (containing drug or placebo).

3. Endogenous Amylin, Related Peptides and Amylin Receptors

Human amylin, a 37-residue polypeptide hormone, is secreted along with insulin from pancreatic β-cells. Loss of β-cell function, which occurs early in type 1 diabetes and late in type 2 diabetes, results in a deficiency of insulin and amylin secretion. Amylin is believed to play a role in regulating glycemia by slowing gastric emptying and promoting satiety, thereby preventing peak postprandial blood glucose levels. The overall effect is to slow the rate of glucose appearance in the blood after feeding.

The amino acid sequence of amylin is most closely related to the sequence of calcitonin gene-related peptide (CGRP). CGRP also shares a similarly positioned disulfide bond and an amidated C-terminus. This is also the case for calcitonin, adrenomedullin and adrenomedullin 2. Taken together, these peptides form a small family united by these characteristic features. Consequently, there is a degree of overlap and pharmacological activity in binding to the cognate receptor for each peptide. The table in FIG. 1 depicts comparative sequence alignments for amylin and certain related reference polypeptides.

Peptides typically designated as members of the calcitonin (CT) peptide family include; calcitonin gene-related peptide (CGRP), calcitonin (CT), amylin (AMY), adrenomedullin 1 and adrenomedullin 2/intermedin (ADM1, ADM2, respectively). Two G protein-coupled receptor proteins (calcitonin receptor; CTR and calcitonin-receptor-like receptor; CALCRL) and three 　receptor activity-modifying proteins (RAMP1, RAMP2, RAMP3) comprise the entire peptide family (CTR, AMY1, AMY2, constituting pharmacologically distinct receptors for AMY3, CGRPR, AM1, AM2). There appear to be at least five distinct receptors (AMY1, AMY2, AMY3, CTR, CGRPR) to which amylin binds with significant affinity. CTR dimerizes with RAMP 1, 2 or 3 to reconstitute AMY1, AMY2 or AMY3 receptors that are pharmacologically selective for amylin over calcitonin. In the absence of RAMP, CTR pharmacology becomes calcitoin selective over amylin. CALCRL dimerized with RAMP1 generates CGRPR with high affinity for CGRP and reduced affinity for all other peptide family members, including amylin. CALCRL and RAMP2 or RAMP3 reconstitute the pharmacology of AM1 and AM2, respectively, with very low or no affinity for amylin.

Amylin analog polypeptides have been developed that have binding affinity for the amylin receptor complex. Pramlintide is a synthetic analogue of human amylin for the treatment of type 1 and type 2 diabetes that uses dietary insulin but cannot achieve desired glycemic control despite optimal insulin therapy, e.g., Amylin Pharmaceuticals. Developed by and approved by the US Food and Drug Administration (FDA). Pramlintide is an amylinmimetic agent that is at least as potent as human amylin. It is also a 37-amino acid polypeptide and differs from the amino acid sequence from human amylin by the replacement of amino acids with proline at positions 25 (alanine), 28 (serine) and 29 (serine). As a result of substitution, pramlintide is soluble, non-adhesive and non-aggregating, thereby overcoming many of the physicochemical obligations of native human amylin. The half-life of pramlintide is approximately 48 minutes in humans, which is longer than native human amylin (approximately 13 minutes). Pramlintide requires frequent and inconvenient administration.

For the treatment of type 1 diabetes, pramlintide is administered up to 4 times per day by subcutaneous injection in the thigh or abdomen before meals as an adjunct to insulin therapy administered after meals. Pramlintide is immiscible with insulin; A separate syringe is used. Pramlintide is administered with or before each meal or snack consisting of at least 250 calories or 30 grams of carbohydrate. A typical starting dose for type 1 diabetes is 15 μg subcutaneous pramlintide before each meal, with subsequent titration up to a target dose of 60 μg before each meal. Reported side effects of pramlintide include nausea and vomiting. In particular, adverse reactions to type 1 diabetes may include severe hypoglycemia. Consequently, the dosage of dietary insulin is reduced for diabetic patients who initiate administration of pramlintide.

For the treatment of type 2 diabetes, pramlintide is administered by subcutaneous injection at a target maintenance dose of 120 μg before each meal with a recommended starting dose of 60 μg.

Davalintide (AC2307) is another analogue of human amylin. Davalintide is a compound under investigation with a half-life of approximately 26 minutes. Like pramlintide, davalintide likewise requires frequent administration by injection.

Certain disclosed amylin analog polypeptides exhibit one or more of good solubility, stability, biological activity and specificity, and longer half-lives for endogenous human amylin and known synthetic amylin analog polypeptides, including Table 3 below. . Certain disclosed amylin analog polypeptides have been developed to accommodate less frequent administration than is necessary for pramlintide. Certain disclosed amylin analog polypeptides have been developed for administration via weekly or monthly injections. Certain disclosed amylin analog polypeptides have been developed for administration via implantation of a delivery device comprising an amylin analog polypeptide, wherein the delivery device is up to 3 months, 6 months, 9 months, 1 year, 18 months or 2 years. amylin analog polypeptide dose of

4. Description of Exemplary Embodiments

In certain embodiments, the present invention relates to an isolated polypeptide that is an amylin analog.

In some embodiments, an isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO:199, or a pharmaceutically acceptable salt thereof:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TX ₃₇ -(OH/NH ₂ )(SEQ ID NO: 199), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H or I;

X ₃ is N or S;

X ₅ is S or A;

X ₆ is T or S;

X ₈ is A or K;

X ₁₀ is Q or S;

X ₁₂ is L or K;

X ₁₃ is A, S, E or K;

X ₁₄ is N, n, d, Y or Q;

X ₁₅ is E, F, f, Y, I, k, K or α-aminoisobutyric acid (Aib);

X ₁₆ is k, K, L, Aib, N -methyl leucine ( N -MeL) or 1;

X ₁₇ is H, V, Q, R, k, K or Aib;

X ₁₈ is K, H or R;

X ₁₉ is S or Aib;

X ₂₀ is S or Aib;

X ₂₂ is N or E;

X ₂₉ is P, R or K;

X ₃₁ is k, K, N or H;

X ₃₅ is e, E, N, K, G, A, Y or P; And

X ₃₇ is Y or P;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge;

provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:199 include:

In some embodiments, the carboxy terminus X ₃₇ is Y-(NH ₂ ). In some embodiments, X ₃₇ at the carboxy terminus is Y-(OH). In some embodiments, the carboxy terminus X ₃₇ is P-(NH ₂ ). In some embodiments, X ₃₇ at the carboxy terminus is P-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k. In some embodiments, X ₁ is H. In some embodiments, X ₁ is I.

As used herein, k refers to D -lysine.

In some embodiments, X ₃ is N. In some embodiments, X ₃ is S.

In some embodiments, X ₅ is S. In some embodiments, X ₅ is A.

In some embodiments, X ₆ is T. In some embodiments, X ₆ is S.

In some embodiments, X ₈ is A. In some embodiments, X ₈ is K.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₂ is L. In some embodiments, X ₁₂ is K.

In some embodiments, X ₁₃ is A. In some embodiments, X ₁₃ is S. In some embodiments, X ₁₃ is E. In some embodiments, X ₁₃ is K.

In some embodiments, X ₁₄ is N. In some embodiments, X ₁₄ is n. In some embodiments, X ₁₄ is d. In some embodiments, X ₁₄ is Y. In some embodiments, X ₁₄ is Q.

As used herein, n refers to D -asparagine.

As used herein, d refers to D -aspartic acid.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F. In some embodiments, X ₁₅ is f. In some embodiments, X ₁₅ is Y. In some embodiments, X ₁₅ is I. In some embodiments, X ₁₅ is K. In some embodiments, X ₁₅ is k. In some embodiments, X ₁₅ is Aib.

As used herein, f refers to D -phenylalanine.

As used herein, Aib alternatively refers to 2-aminoisobutyric acid, α-aminoisobutyric acid, α-methylalanine or 2-methylalanine.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is l. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k. In some embodiments, X ₁₆ is Aib. In some embodiments, X ₁₆ is N -MeL.

As used herein, l refers to D -leucine.

As used herein, N- MeL refers to N -methyl leucine.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q. In some embodiments, X ₁₇ is R. In some embodiments, X ₁₇ is K. In some embodiments, X ₁₇ is k. In some embodiments, X ₁₇ is Aib.

In some embodiments, X ₁₈ is K. In some embodiments, X ₁₈ is H. In some embodiments, X ₁₈ is R.

In some embodiments, X ₁₉ is S. In some embodiments, X ₁₉ is Aib.

In some embodiments, X ₂₀ is S. In some embodiments, X ₂₀ is Aib.

In some embodiments, X ₂₂ is N. In some embodiments, X ₂₂ is E.

In some embodiments, X ₂₉ is P. In some embodiments, X ₂₉ is R. In some embodiments, X ₂₉ is K.

In some embodiments, X ₃₁ is k. In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N. In some embodiments, X ₃₁ is H.

In some embodiments, X ₃₅ is e. In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N. In some embodiments, X ₃₅ is K. In some embodiments, X ₃₅ is G. In some embodiments, X ₃₅ is A. In some embodiments, X ₃₅ is Y. In some embodiments, X ₃₅ is P.

As used herein, e refers to D -glutamic acid.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:199 include:

In some embodiments, X ₁ is S and X ₅ is S. In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₈ is K. In some embodiments, X ₁ is S and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E. In some embodiments, X ₁ is S and X ₃₇ is Y.

In some embodiments, X ₁ is K and X ₅ is S. In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₈ is K. In some embodiments, X ₁ is K and X ₃₁ is K. In some embodiments, X ₁ is K and X ₃₅ is E. In some embodiments, X ₁ is K and X ₃₇ is Y.

In some embodiments, X ₁ is k and X ₅ is S. In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₈ is K. In some embodiments, X ₁ is k and X ₃₁ is K. In some embodiments, X ₁ is k and X ₃₅ is E. In some embodiments, X ₁ is k and X ₃₇ is Y.

In some embodiments, X ₅ is S and X ₁₅ is E. In some embodiments, X ₅ is S and X ₁₀ is Q. In some embodiments, X ₅ is S and X ₁₆ is L. In some embodiments, X ₅ is S and X ₁₆ is k. In some embodiments, X ₅ is S and X ₁₇ is H. In some embodiments, X ₅ is S and X ₁₈ is K. In some embodiments, X ₅ is S and X ₃₁ is K. In some embodiments, X ₅ is S and X ₃₅ is E. In some embodiments, X ₅ is S and X ₃₇ is Y.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₈ is K. In some embodiments, X ₁₀ is Q and X ₃₁ is K. In some embodiments, X ₁₀ is Q and X ₃₅ is E. In some embodiments, X ₁₀ is Q and X ₃₇ is Y.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₈ is K. In some embodiments, X ₁₅ is E and X ₃₁ is K. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₇ is Y.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₈ is K. In some embodiments, X ₁₆ is L and X ₃₁ is K. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₇ is Y.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₈ is K. In some embodiments, X ₁₆ is k and X ₃₁ is K. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₇ is Y.

In some embodiments, X ₁₇ is H and X ₁₈ is K. In some embodiments, X ₁₇ is H and X ₃₁ is K. In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₇ is Y.

In some embodiments, X ₁₈ is K and X ₃₁ is K. In some embodiments, X ₁₈ is K and X ₃₅ is E. In some embodiments, X ₁₈ is K and X ₃₇ is Y.

In some embodiments, X ₃₁ is K and X ₃₅ is E. In some embodiments, X ₃₁ is K and X ₃₇ is Y.

In some embodiments, X ₃₅ is E and X ₃₇ is Y.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:199 include:

In some embodiments, X ₁ is S, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₇ is Y.

In some embodiments, X ₁ is K, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₇ is Y.

In some embodiments, X ₁ is k, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₅ is E. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₇ is Y.

In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is k. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₇ is H. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₈ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₁ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₅ is E. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₇ is Y.

In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₁₀ is Q, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₀ is Q, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₀ is Q, X ₃₁ is K, and X ₃₅ is E. In some embodiments, X ₁₀ is Q, X ₃₁ is K, and X ₃₇ is Y.

In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₅ is E. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₇ is Y.

In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₅ is E. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₇ is Y.

In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₅ is E. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₇ is Y.

In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₅ is E. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₇ is Y.

In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₅ is E. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₇ is Y.

In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₅ is E. In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₇ is Y.

In some embodiments, X ₃₁ is K, X ₃₅ is E, and X ₃₇ is Y.

In some embodiments, the carboxy terminus amino acid 37 is Y-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is Y-(OH). In some embodiments, the carboxy terminus amino acid 37 is P-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is P-(OH).

In some embodiments, an isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO:200, or a pharmaceutically acceptable salt thereof:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200 ), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H or I;

X ₃ is N or S;

X ₅ is S or A;

X ₆ is T or S;

X ₈ is A or K;

X ₁₀ is Q or S;

X ₁₂ is L or K;

X ₁₃ is A, S, E or K;

X ₁₄ is N, n, d, Y or Q;

X ₁₅ is E, F, f, Y, I, k, K or α-aminoisobutyric acid (Aib);

X ₁₆ is k, K, L, Aib, N -methyl leucine ( N -MeL) or 1;

X ₁₇ is H, V, Q, R, k, K or Aib;

X ₁₈ is K, H or R;

X ₁₉ is S or Aib;

X ₂₀ is S or Aib;

X ₂₂ is N or E;

X ₂₉ is P, R or K;

X ₃₁ is k, K or N; And

X ₃₅ is e, E or N;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge;

provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, an isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 201, or a pharmaceutically acceptable salt thereof: X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 201), wherein:

X ₁ is S, K, k, H or I;

X ₃ is N or S;

X ₅ is S or A;

X ₆ is T or S;

X ₈ is A or K;

X ₁₀ is Q or S;

X ₁₂ is L or K;

X ₁₃ is A, S, E or K;

X ₁₄ is N, n, d, Y or Q;

X ₁₅ is E, F, f, Y, I, k, K or Aib;

X ₁₆ is k, K, L, Aib, N -MeL or 1;

X ₁₇ is H, V, Q, R, k, K or Aib;

X ₁₈ is K, H or R;

X ₁₉ is S or Aib;

X ₂₀ is S or Aib;

X ₂₂ is N or E; And

X ₂₉ is P, R or K;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

The _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:200 or SEQ ID NO:201 include:

In some embodiments, the carboxy terminus amino acid 37 is Y-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k. In some embodiments, X ₁ is H. In some embodiments, X ₁ is I.

As used herein, k refers to D -lysine.

In some embodiments, X ₃ is N. In some embodiments, X ₃ is S.

In some embodiments, X ₅ is S. In some embodiments, X ₅ is A.

In some embodiments, X ₆ is T. In some embodiments, X ₆ is S.

In some embodiments, X ₈ is A. In some embodiments, X ₈ is K.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₂ is L. In some embodiments, X ₁₂ is K.

In some embodiments, X ₁₃ is A. In some embodiments, X ₁₃ is S. In some embodiments, X ₁₃ is E. In some embodiments, X ₁₃ is K.

In some embodiments, X ₁₄ is N. In some embodiments, X ₁₄ is n. In some embodiments, X ₁₄ is d. In some embodiments, X ₁₄ is Y. In some embodiments, X ₁₄ is Q.

As used herein, n refers to D -asparagine.

As used herein, d refers to D -aspartic acid.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F. In some embodiments, X ₁₅ is f. In some embodiments, X ₁₅ is Y. In some embodiments, X ₁₅ is I. In some embodiments, X ₁₅ is K. In some embodiments, X ₁₅ is k. In some embodiments, X ₁₅ is Aib.

As used herein, f refers to D -phenylalanine.

As used herein, Aib alternatively refers to 2-aminoisobutyric acid, α-aminoisobutyric acid, α-methylalanine or 2-methylalanine.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is l. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k. In some embodiments, X ₁₆ is Aib. In some embodiments, X ₁₆ is N -MeL.

As used herein, l refers to D -leucine.

As used herein, N- MeL refers to N -methyl leucine.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q. In some embodiments, X ₁₇ is R. In some embodiments, X ₁₇ is K. In some embodiments, X ₁₇ is k. In some embodiments, X ₁₇ is Aib.

In some embodiments, X ₁₈ is K. In some embodiments, X ₁₈ is H. In some embodiments, X ₁₈ is R.

In some embodiments, X ₁₉ is S. In some embodiments, X ₁₉ is Aib.

In some embodiments, X ₂₀ is S. In some embodiments, X ₂₀ is Aib.

In some embodiments, X ₂₂ is N. In some embodiments, X ₂₂ is E.

In some embodiments, X ₂₉ is P. In some embodiments, X ₂₉ is R. In some embodiments, X ₂₉ is K.

In some embodiments, X ₃₁ is k. In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is e. In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

As used herein, e refers to D -glutamic acid.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:200 or SEQ ID NO:201 include:

In some embodiments, X ₁ is S and X ₅ is S. In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₈ is K. In some embodiments, X ₁ is S and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E.

In some embodiments, X ₁ is K and X ₅ is S. In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₈ is K. In some embodiments, X ₁ is K and X ₃₁ is K. In some embodiments, X ₁ is K and X ₃₅ is E.

In some embodiments, X ₁ is k and X ₅ is S. In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₈ is K. In some embodiments, X ₁ is k and X ₃₁ is K. In some embodiments, X ₁ is k and X ₃₅ is E.

In some embodiments, X ₅ is S and X ₁₅ is E. In some embodiments, X ₅ is S and X ₁₀ is Q. In some embodiments, X ₅ is S and X ₁₆ is L. In some embodiments, X ₅ is S and X ₁₆ is k. In some embodiments, X ₅ is S and X ₁₇ is H. In some embodiments, X ₅ is S and X ₁₈ is K. In some embodiments, X ₅ is S and X ₃₁ is K. In some embodiments, X ₅ is S and X ₃₅ is E.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₈ is K. In some embodiments, X ₁₀ is Q and X ₃₁ is K. In some embodiments, X ₁₀ is Q and X ₃₅ is E.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₈ is K. In some embodiments, X ₁₅ is E and X ₃₁ is K. In some embodiments, X ₁₅ is E and X ₃₅ is E.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₈ is K. In some embodiments, X ₁₆ is L and X ₃₁ is K. In some embodiments, X ₁₆ is L and X ₃₅ is E.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₈ is K. In some embodiments, X ₁₆ is k and X ₃₁ is K. In some embodiments, X ₁₆ is k and X ₃₅ is E.

In some embodiments, X ₁₇ is H and X ₁₈ is K. In some embodiments, X ₁₇ is H and X ₃₁ is K. In some embodiments, X ₁₇ is H and X ₃₅ is E.

In some embodiments, X ₁₈ is K and X ₃₁ is K. In some embodiments, X ₁₈ is K and X ₃₅ is E.

In some embodiments, X ₃₁ is K and X ₃₅ is E.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:200 or SEQ ID NO:201 include:

In some embodiments, X ₁ is S, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E.

In some embodiments, X ₁ is K, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₁ is k, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is k. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₇ is H. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₈ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₁ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₅ is E.

In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₁₀ is Q, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₀ is Q, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₀ is Q, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₅ is E.

In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₅ is E.

In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, the isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 202, or a pharmaceutically acceptable salt thereof: X ₁ CNTX ₅ TCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ X ₁₈ SSNNFGPILPPTX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 202), wherein:

X ₁ is S, k or K;

X ₅ is S or A;

X ₁₀ is Q or S;

X ₁₅ is E or F;

X ₁₆ is k, K or L;

X ₁₇ is H, V and Q;

X ₁₈ is K, H or R;

X ₃₁ is K or N; And

X ₃₅ is E or N;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

two cysteine residues of X ₁ CNTX ₅ TC (SEQ ID NO: 308) are optionally further linked by a disulfide bridge;

provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 203, or a pharmaceutically acceptable salt thereof: X ₁ CNTX ₅ TCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ X ₁₈ SSNNFGPILPPTKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 203), wherein:

X ₁ is S, k or K;

X ₅ is S or A;

X ₁₀ is Q or S;

X ₁₅ is E or F;

X ₁₆ is k, K or L;

X ₁₇ is H, V and Q; And

X ₁₈ is K, H or R;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

The _{two cysteine residues of X 1} CNTX ₅ TC (SEQ ID NO: 308) are optionally further linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:202 or SEQ ID NO:203 include:

In some embodiments, the carboxy terminus amino acid 37 is Y-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k.

In some embodiments, X ₅ is S. In some embodiments, X ₅ is A.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₁₈ is K. In some embodiments, X ₁₈ is H. In some embodiments, X ₁₈ is R.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:202 or SEQ ID NO:203 include:

In some embodiments, X ₁ is S and X ₅ is S. In some embodiments, X ₁ is S and X ₁₀ is E. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₈ is K. In some embodiments, X ₁ is S and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E.

In some embodiments, X ₁ is K and X ₅ is S. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₈ is K. In some embodiments, X ₁ is K and X ₃₁ is K. In some embodiments, X ₁ is K and X ₃₅ is E.

In some embodiments, X ₅ is S and X ₁₅ is E. In some embodiments, X ₅ is S and X ₁₆ is L. In some embodiments, X ₅ is S and X ₁₆ is k. In some embodiments, X ₅ is S and X ₁₇ is H. In some embodiments, X ₅ is S and X ₁₈ is K. In some embodiments, X ₅ is S and X ₃₁ is K. In some embodiments, X ₅ is S and X ₃₅ is E.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₈ is K. In some embodiments, X ₁₀ is Q and X ₃₁ is K. In some embodiments, X ₁₀ is Q and X ₃₅ is E.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₈ is K. In some embodiments, X ₁₅ is E and X ₃₁ is K. In some embodiments, X ₁₅ is E and X ₃₅ is E.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₈ is K. In some embodiments, X ₁₆ is L and X ₃₁ is K. In some embodiments, X ₁₆ is L and X ₃₅ is E.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₈ is K. In some embodiments, X ₁₆ is k and X ₃₁ is K. In some embodiments, X ₁₆ is k and X ₃₅ is E.

In some embodiments, X ₁₇ is H and X ₁₈ is K. In some embodiments, X ₁₇ is H and X ₃₁ is K. In some embodiments, X ₁₇ is H and X ₃₅ is E.

In some embodiments, X ₁₈ is K and X ₃₁ is K. In some embodiments, X ₁₈ is K and X ₃₅ is E.

In some embodiments, X ₃₁ is K and X ₃₅ is E.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:202 or SEQ ID NO:203 include:

In some embodiments, X ₁ is S, X ₅ is S, and X ₁₀ is Q. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E.

In some embodiments, X ₁ is K, X ₅ is S, and X ₁₀ is Q. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₅ is E. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₆ is L. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₆ is k. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₇ is H. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₈ is K. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₃₁ is K. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₃₅ is E.

In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₆ is k. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₇ is H. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₈ is K. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₃₁ is K. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₅ is E.

In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₅ is E.

In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, the isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 204), wherein:

X ₁ is S, K or k;

X ₁₀ is Q or S;

X ₁₅ is E or F;

X ₁₆ is L, K or k;

X ₁₇ is H, V or Q; And

X ₃₅ is E or N;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

The _{two cysteine residues of X 1} CNTSTC (SEQ ID NO: 309) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 205, or a pharmaceutically acceptable salt thereof: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 205), wherein:

X ₁ is S, K or k;

X ₁₀ is Q or S;

X ₁₅ is E or F;

X ₁₆ is L, K or k; And

X ₁₇ is H, V or Q;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; And

The _{two cysteine residues of X 1} CNTSTC (SEQ ID NO: 309) are optionally further linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:204 or SEQ ID NO:205 include:

In some embodiments, the carboxy terminus amino acid 37 is Y-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:204 or SEQ ID NO:205 include:

In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₀ is S. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₅ is F. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is K. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₇ is V. In some embodiments, X ₁ is S and X ₁₇ is Q. In some embodiments, X ₁ is S and X ₃₅ is E. In some embodiments, X ₁ is S and X ₃₅ is N.

In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₀ is S. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₅ is F. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is K. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₇ is V. In some embodiments, X ₁ is K and X ₁₇ is Q. In some embodiments, X ₁ is K and X ₃₅ is E. In some embodiments, X ₁ is K and X ₃₅ is N.

In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₀ is S. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₅ is F. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is K. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₇ is V. In some embodiments, X ₁ is k and X ₁₇ is Q. In some embodiments, X ₁ is k and X ₃₅ is E. In some embodiments, X ₁ is k and X ₃₅ is N.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₅ is F. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is K. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₇ is V. In some embodiments, X ₁₀ is Q and X ₁₇ is Q. In some embodiments, X ₁₀ is Q and X ₃₅ is E. In some embodiments, X ₁₀ is Q and X ₃₅ is N.

In some embodiments, X ₁₀ is S and X ₁₅ is E. In some embodiments, X ₁₀ is S and X ₁₅ is F. In some embodiments, X ₁₀ is S and X ₁₆ is L. In some embodiments, X ₁₀ is S and X ₁₆ is K. In some embodiments, X ₁₀ is S and X ₁₆ is k. In some embodiments, X ₁₀ is S and X ₁₇ is H. In some embodiments, X ₁₀ is S and X ₁₇ is V. In some embodiments, X ₁₀ is S and X ₁₇ is Q. In some embodiments, X ₁₀ is S and X ₃₅ is E. In some embodiments, X ₁₀ is S and X ₃₅ is N.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₅ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₇ is V. In some embodiments, X ₁₆ is k and X ₁₇ is Q. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, the isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 206, or a pharmaceutically acceptable salt thereof: SCNTSTCATQRLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 206), wherein:

X ₁₅ is E or F;

X ₁₆ is L, K or k;

X ₁₇ is H, V or Q; And

X ₃₅ is E or N;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; And

The two cysteine residues of SCNTSTC (SEQ ID NO: 310) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 207, or a pharmaceutically acceptable salt thereof: SCNTSTCATQRLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 207), wherein:

X ₁₅ is E or F;

X ₁₆ is L, K or k; And

X ₁₇ is H, V or Q;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; And

The two cysteine residues of SC*NTSTC* are optionally further linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:206 or SEQ ID NO:207 include:

In some embodiments, the carboxy terminus amino acid 37 is Y-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is Y-(OH).

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₅ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₇ is V. In some embodiments, X ₁₆ is k and X ₁₇ is Q. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, an isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of:

SC*NTSTC*ATQRLANFkHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 127), also referred to herein as Compound A127;

_{SC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH 2} ) (SEQ ID NO: 57), also referred to herein as compound compound A57;

_{SC*NTSTC*ATQRLANEKHKSSNNFGPILPPTKVGSETY-(NH 2} ) (SEQ ID NO: 128), also referred to herein as compound compound A128;

SC*NTSTC*ATQRLANEkHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 129), also referred to herein as Compound A129; and

_{SC*NTSTC*ATQRLANFLVKSSNEFGPILPPTKVGSETY-(NH 2} ) (SEQ ID NO: 43), also referred to herein as compound compound A43.

In some embodiments, an isolated polypeptide of the present disclosure comprises the following amino acid sequence: SC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 57), also referred to herein as Compound A57.

In some embodiments, an isolated polypeptide of the present disclosure comprises the following amino acid sequence: SC*NTSTC*ATQRLANFkHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 127), also referred to herein as Compound A127.

In some embodiments, an isolated polypeptide of the present disclosure comprises the following amino acid sequence: SC*NTSTC*ATQRLANEk*((γGlu) ₂ -CO(CH ₂ ) ₁₄ CH _{3 , also referred to herein as Compound A27} ) HKSSNNFGPILPP TKVGSETY-NH ₂ (SEQ ID NO: 27).

In some embodiments, the isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 208), wherein:

X ₁ is K or k;

X ₁₀ is Q or S;

X ₁₅ is E or F;

X ₁₆ is L, K or k;

X ₁₇ is H, V or Q; And

X ₃₅ is E or N;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; And

The _{two cysteine residues of X 1} CNTSTC (SEQ ID NO: 318) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 209, or a pharmaceutically acceptable salt thereof: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 209), wherein:

X ₁ is K or k;

X ₁₀ is Q or S;

X ₁₅ is E or F;

X ₁₆ is L, K or k; And

X ₁₇ is H, V or Q;

each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; And

The _{two cysteine residues of X 1} CNTSTC (SEQ ID NO: 318) are optionally further linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO:208 or SEQ ID NO:209 include:

In some embodiments, the carboxy terminus amino acid 37 is Y-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is Y-(OH).

In some embodiments, X ₁ is K. In some embodiments, X ₁ is k.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₀ is S. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₅ is F. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is K. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₇ is V. In some embodiments, X ₁ is K and X ₁₇ is Q. In some embodiments, X ₁ is K and X ₃₅ is E. In some embodiments, X ₁ is K and X ₃₅ is N.

In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₀ is S. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₅ is F. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is K. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₇ is V. In some embodiments, X ₁ is k and X ₁₇ is Q. In some embodiments, X ₁ is k and X ₃₅ is E. In some embodiments, X ₁ is k and X ₃₅ is N.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₅ is F. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is K. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₇ is V. In some embodiments, X ₁₀ is Q and X ₁₇ is Q. In some embodiments, X ₁₀ is Q and X ₃₅ is E. In some embodiments, X ₁₀ is Q and X ₃₅ is N.

In some embodiments, X ₁₀ is S and X ₁₅ is E. In some embodiments, X ₁₀ is S and X ₁₅ is F. In some embodiments, X ₁₀ is S and X ₁₆ is L. In some embodiments, X ₁₀ is S and X ₁₆ is K. In some embodiments, X ₁₀ is S and X ₁₆ is k. In some embodiments, X ₁₀ is S and X ₁₇ is H. In some embodiments, X ₁₀ is S and X ₁₇ is V. In some embodiments, X ₁₀ is S and X ₁₇ is Q. In some embodiments, X ₁₀ is S and X ₃₅ is E. In some embodiments, X ₁₀ is S and X ₃₅ is N.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₅ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, an isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of:

_{KC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH 2} ) (SEQ ID NO: 130), also referred to herein as Compound A130; and

KC*NTSTC*ATQRLANFLQKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 131), also referred to herein as compound A131.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

5. Conjugation of a lipophilic substituent to any peptide optionally via a spacer

In some embodiments, any of the disclosed polypeptides are each optionally substituted with one or more lipophilic substituents, optionally via a spacer, wherein “lipophilic substituents” and “spacers” are defined herein. In some embodiments, any disclosed polypeptide comprising an amino acid sequence selected from the group consisting of the amino acid sequence represented by any consensus sequence of SEQ ID NOs: 1-143, each optionally comprising one or more lipophilic substituents via a spacer , may be modified, or each optionally further modified by covalent attachment of one or more lipophilic substituents via a spacer. In some embodiments, the lipophilic substituent may be attached to an amino group of the polypeptide (eg, the ε-amino group of a lysine residue) by a carboxyl group of the lipophilic substituent, or optionally an amino group of the spacer, wherein the carboxyl group of the spacer It forms an amide bond with the ε-amino group of the lysine residue.

lipophilic substituents

Conjugation of one or more "lipophilic substituents" to any of the disclosed polypeptides, each optionally via a "spacer", is intended to prolong polypeptide action by facilitating binding to serum albumin, wherein the conjugated poly Delays renal clearance of peptides. As used herein, “lipophilic substituent” includes substituents containing from 4 to 40 carbon atoms, from 8 to 25 carbon atoms, from 12 to 22 carbon atoms, or from 6 to 20 carbon atoms. The lipophilic substituent may be attached to an amino group of the polypeptide (eg, the ε-amino group of a lysine residue) by the carboxyl group of the lipophilic substituent, or optionally the amino group of the spacer, the carboxyl group of the spacer in turn being the amino acid to which it is attached. Forms an amide bond with the amino group of a residue (eg, lysine). In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituent with or without optional spacers as defined in more detail below.

In some embodiments, the lipophilic substituent comprises a straight-chain or branched alkyl group. In some embodiments, the lipophilic substituent is an acyl group of a straight-chain or branched fatty acid. In some embodiments, the lipophilic substituent is an acyl group of a straight-chain or branched fatty acid, which is further substituted with one or more carboxylic acid and/or hydroxamic acid groups.

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each without optional spacers. In some embodiments, the lipophilic substituent is a monovalent group of Formula I:

during the meal,

Z is —CH ₃ or —CO ₂ H; And

m is 4 to 24,

The lipophilic substituent then forms an amide bond between the amino group of the disclosed polypeptide (eg, the ε-amino group of lysine) and the CO- group of the lipophilic substituent.

In some embodiments, m is selected from the group consisting of 4-20, 8-20, 12-20, 14-20, 16-20, 14, 16, 18 and 20.

In some embodiments, Z is —CO ₂ H and the lipophilic substituent has the formula —CO—(CH ₂ ) _m —CO ₂ H. In some embodiments, -CO-(CH ₂ ) _m -Z is -CO-(CH ₂ ) ₄ -CO ₂ H, -CO-(CH ₂ ) ₅ -CO ₂ H, -CO-(CH ₂ ) ₆ -CO ₂ H, -CO-(CH ₂ ) ₇ -CO ₂ H, -CO-(CH ₂ ) ₈ -CO ₂ H, -CO-(CH ₂ ) ₉ -CO ₂ H, -CO-(CH _{2 )} ) ₁₀ -CO ₂ H, -CO-(CH ₂ ) ₁₁ -CO ₂ H, -CO-(CH ₂ ) ₁₂ -CO ₂ H, -CO-(CH ₂ ) ₁₃ -CO ₂ H, -CO-( CH ₂ ) ₁₄ -CO ₂ H, -CO-(CH ₂ ) ₁₅ -CO ₂ H, -CO-(CH ₂ ) ₁₆ -CO ₂ H, -CO-(CH ₂ ) ₁₇ -CO ₂ H, -CO -(CH ₂ ) ₁₈ -CO ₂ H, -CO-(CH ₂ ) ₁₉ -CO ₂ H, -CO-(CH ₂ ) ₂₀ -CO ₂ H is selected from the group consisting of.

In some embodiments, the lipophilic substituent is —CO—(CH ₂ ) ₁₈ —CO ₂ H.

In some embodiments, Z is —CH ₃ and the lipophilic substituent has the formula —CO—(CH ₂ ) _m —CH ₃ . In some embodiments, -CO-(CH ₂ ) _m -Z is -CO-(CH ₂ ) ₄ -CH ₃ , -CO-(CH ₂ ) ₅ -CH ₃ , -CO-(CH ₂ ) ₆ -CH ₃ , -CO-(CH ₂ ) ₇ -CH ₃ , -CO-(CH ₂ ) ₈ -CH ₃ , -CO-(CH ₂ ) ₉ -CH ₃ , -CO-(CH ₂ ) ₁₀ -CH ₃ , -CO-(CH ₂ ) ₁₁ -CH ₃ , -CO-(CH ₂ ) ₁₂ -CH ₃ , -CO-(CH ₂ ) ₁₃ -CH ₃ , -CO-(CH ₂ ) ₁₄ -CH ₃ , -CO -(CH ₂ ) ₁₅ -CH ₃ , -CO-(CH ₂ ) ₁₆ -CH ₃ , -CO-(CH ₂ ) ₁₇ -CH ₃ , -CO-(CH ₂ ) ₁₈ -CH ₃ , -CO-( CH ₂ ) ₁₉ -CH ₃ and -CO-(CH ₂ ) ₂₀ -CH ₃ .

spacer

In some embodiments, the lipophilic substituent is attached to the parent peptide by a “spacer”. In some embodiments, provided herein is any disclosed polypeptide comprising an amino acid sequence selected from the group consisting of the amino acid sequence represented by any consensus sequence of SEQ ID NO: 1 to SEQ ID NO: 143, comprising a lipophilic substituent herein However, the lipophilic substituent is linked to the ε-amino group of lysine through a spacer, which spacer forms a bridge between the amino group of the disclosed polypeptide and the CO-group of the lipophilic substituent.

In some embodiments, the spacer comprises one or more amino acids, e.g., a single amino acid, e.g., Glu, Asp, Gly or Lys, a dipeptide such as 2(Glu), Glu-Gly, or a polypeptide, e.g., 3( Glu), 4(Glu) (SEQ ID NO: 317), 2(Glu)-Gly, and the like. In some embodiments, when the spacer comprises one or more amino acids, e.g., Glu, Asp, Gly or Lys, one carboxyl group of the spacer is capable of forming an amide bond with an amino group of the disclosed polypeptide and the amino group of the spacer can form an amide bond with the carboxyl group of the lipophilic substituent.

In some embodiments, when the spacer comprises Glu or Asp, it further comprises a carboxylic acid-terminal side chain, wherein the terminal carboxyl group of the Glu or Asp-containing spacer side chain is capable of forming an amide bond with an amino group of a disclosed polypeptide; , Glu or the amino group of the Asp-containing spacer can form an amide bond with the carboxyl group of a lipophilic substituent, ie, γGlu or βAsp. In some embodiments, the spacer is γGlu. In some embodiments, the spacer is 2(γGlu). In some embodiments, the spacer is 3(γGlu).

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each with a spacer. In some embodiments, the lipophilic substituent and spacer are monovalent groups of Formula II:

during the meal,

Y is selected from the group consisting of γGlu, Asp, Lys and Gly;

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24; And

n is 1 to 10.

In some embodiments, Y is selected from the group consisting of γGlu and Gly. In some embodiments, Y is γGlu. In some embodiments, Y is Gly.

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each with a spacer. In some embodiments, the lipophilic substituent and spacer are monovalent groups of Formula III:

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24; And

n is 1 to 10.

In some embodiments, Z is —CH ₃ . In some embodiments, Z is —CO ₂ H.

In some embodiments, m is selected from the group consisting of 4-20, 8-20, 12-20, 14-20, 16-20, 14, 16, 18 and 20.

In some embodiments, n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each with a spacer. In some embodiments, the lipophilic substituent and spacer are monovalent groups of Formula IV:

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24; And

n is 1 to 10.

In some embodiments, (γGlu) _n is γGlu; 2(γGlu); 3(γGlu); 4(γGlu) (SEQ ID NO: 313); and 5(γGlu) (SEQ ID NO: 314). In some embodiments, -(γGlu) _n -(Gly)-(“(γGlu) _n -(Gly)” set forth in SEQ ID NO: 312) is 2(γGlu),Gly; and 3 (γGlu), Gly (SEQ ID NO: 315).

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each with a spacer. In some embodiments, the lipophilic substituent and spacer are monovalent groups of Formula (V):

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24; And

n is 1 to 10.

In some embodiments, certain variables represented by Formula II, Formula III, Formula IV, or Formula V include:

In some embodiments, Z is —CH ₃ . In some embodiments, Z is —CO ₂ H.

In some embodiments, m is selected from the group consisting of 4-20, 8-20, 12-20, 14-20, 16-20, 14, 16, 18 and 20.

In some embodiments, n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

In some embodiments, n is 1 and Z is —CO ₂ H. In some embodiments, n is 1 and Z is —CH ₃ . In some embodiments, n is 2 and Z is —CO ₂ H. In some embodiments, n is 2 and Z is —CH ₃ . In some embodiments, n is 3 and Z is —CO ₂ H. In some embodiments, n is 3 and Z is —CH ₃ . In some embodiments, n is 4 and Z is —CO ₂ H. In some embodiments, n is 4 and Z is —CH ₃ . In some embodiments, n is 5 and Z is —CO ₂ H. In some embodiments, n is 5 and Z is —CH ₃ .

In some embodiments, n is 1, Z is —CO ₂ H, and m is 14-20. In some embodiments, n is 1, Z is —CO ₂ H, and m is 14. In some embodiments, n is 1, Z is —CO ₂ H, and m is 16. In some embodiments, n is 1, Z is —CO ₂ H, and m is 18.

In some embodiments, n is 1, Z is —CH ₃ , and m is 14-20. In some embodiments, n is 1, Z is —CH ₃ , and m is 14. In some embodiments, n is 1, Z is —CH ₃ , and m is 16. In some embodiments, n is 1, Z is —CH ₃ , and m is 18.

In some embodiments, n is 2, Z is —CO ₂ H, and m is 14-20. In some embodiments, n is 2, Z is —CO ₂ H, and m is 14. In some embodiments, n is 2, Z is —CO ₂ H, and m is 16. In some embodiments, n is 2, Z is —CO ₂ H, and m is 18.

In some embodiments, n is 2, Z is —CH ₃ , and m is 14-20. In some embodiments, n is 2, Z is —CH ₃ , and m is 14. In some embodiments, n is 2, Z is —CH ₃ , and m is 16. In some embodiments, n is 2, Z is —CH ₃ , and m is 18.

In some embodiments, n is 3, Z is —CO ₂ H, and m is 14-20. In some embodiments, n is 3, Z is —CO ₂ H, and m is 14. In some embodiments, n is 3, Z is —CO ₂ H, and m is 16. In some embodiments, n is 3, Z is —CO ₂ H, and m is 18.

In some embodiments, n is 3, Z is —CH ₃ , and m is 14-20. In some embodiments, n is 3, Z is —CH ₃ , and m is 14. In some embodiments, n is 3, Z is —CH ₃ , and m is 16. In some embodiments, n is 3, Z is —CH ₃ , and m is 18.

In some embodiments, n is 4, Z is —CO ₂ H, and m is 14-20. In some embodiments, n is 4, Z is —CO ₂ H, and m is 14. In some embodiments, n is 4, Z is —CO ₂ H, and m is 16. In some embodiments, n is 4, Z is —CO ₂ H, and m is 18.

In some embodiments, n is 4, Z is —CH ₃ , and m is 14-20. In some embodiments, n is 4, Z is —CH ₃ , and m is 14. In some embodiments, n is 4, Z is —CH ₃ , and m is 16. In some embodiments, n is 4, Z is —CH ₃ , and m is 18.

In some embodiments, n is 5, Z is —CO ₂ H, and m is 14-20. In some embodiments, n is 5, Z is —CO ₂ H, and m is 14. In some embodiments, n is 5, Z is —CO ₂ H, and m is 16. In some embodiments, n is 5, Z is —CO ₂ H, and m is 18.

In some embodiments, n is 5, Z is —CH ₃ , and m is 14-20. In some embodiments, n is 5, Z is —CH ₃ , and m is 14. In some embodiments, n is 5, Z is —CH ₃ , and m is 16. In some embodiments, n is 5, Z is —CH ₃ , and m is 18.

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each with a spacer. In some embodiments, the lipophilic substituent and spacer are monovalent groups of Formula VI:

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24;

Y1 is selected from the group consisting of γGlu, Asp and Gly;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

V is -[COCH ₂ (O(CH ₂ ) ₂ ) _t OCH ₂ NH]-, t is 1 to 8;

r is 1 to 8;

n1 is 0 to 10; And

n2 is 0 to 10.

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each with a spacer. In some embodiments, the lipophilic substituent and spacer are monovalent groups of Formula VII:

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24;

Y1 is selected from the group consisting of γGlu, Asp and Gly;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

dpeg is -[CO(CH ₂ )O(CH ₂ ) ₂ O(CH ₂ )NH]-;

r is 1 to 8;

n1 is 0 to 10; And

n2 is 0 to 10.

In some embodiments, -(Y1) _n1 -(dpeg) _r -(Y2) _n2 - is γGlu,dpeg,dpeg,γGlu; γGlu, dpeg, dpeg, 2 (γGlu); γGlu, dpeg, dpeg, 3 (γGlu); γGlu, dpeg, dpeg, 4 (γGlu); 2(γGlu), dpeg, dpeg, γGlu; and 2(γGlu),dpeg,γGlu.

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each with a spacer. In some embodiments, the lipophilic substituent and spacer are monovalent groups of Formula (VIII):

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

V is -[COCH ₂ (O(CH ₂ ) ₂ ) _t OCH ₂ NH]-, t is 1 to 8;

r is 1 to 8; And

n2 is 0 to 10.

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each with a spacer. In some embodiments, the lipophilic substituent and spacer are monovalent groups of Formula (IX):

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24;

dpeg is -[CO(CH ₂ )O(CH ₂ ) ₂ O(CH ₂ )NH]-;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

r is 1 to 8; And

n2 is 0 to 10.

In some embodiments, -(dpeg) _r -(Y2) _n2 - is dpeg,γGlu; and dpeg, dpeg, γGlu.

In some embodiments, the polypeptide comprises 3, 2 or preferably 1 lipophilic substituents each with a spacer. In some embodiments, the lipophilic substituent and spacer are monovalent groups of Formula X:

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24;

Y1 is selected from the group consisting of γGlu, Asp and Gly;

dpeg is -[CO(CH ₂ )O(CH ₂ ) ₂ O(CH ₂ )NH]-;

r is 1 to 8; And

n1 is 0 to 10.

In some embodiments, -(Y1) _n1 -(dpeg) _r - is 2(γGlu),dpeg.

In some embodiments, the spacer comprises a divalent group of Formula (XI):

during the meal,

each of R ₁ and R ₃ is hydrogen or C ₁ -C ₄ alkyl;

each R ₂ is H or CO ₂ H;

p is 1, 2, 3, 4, 5 or 6;

q is 1, 2 or 3;

r is 0 or 1.

This spacer forms a bridge between the amino group of the disclosed polypeptides and the CO- group of the lipophilic substituent.

In some embodiments, the spacer comprises a divalent group of Formula (XII):

during the meal,

each R ₃ is hydrogen or C ₁ -C ₄ alkyl;

q is 1, 2 or 3;

r is 0 or 1.

This spacer forms a bridge between the amino group of the disclosed polypeptides and the CO- group of the lipophilic substituent.

In some embodiments, certain variables represented by Formula XI or Formula XII include:

In some embodiments, each R ₁ is hydrogen. In some embodiments, each R ₃ is hydrogen. In some embodiments, each R ₁ and each R ₃ is hydrogen.

In some embodiments, at least one R ₂ is CO ₂ H. In some embodiments, one R ₂ is CO ₂ H.

In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5. In some embodiments, p is 6.

In some embodiments, q is 1. In some embodiments, q is 2. In some embodiments, q is 3.

In some embodiments, r is 0. In some embodiments, r is 1.

In some embodiments, the spacer is γ-glutamyl, ie, —NH(CHCO ₂ H)(CH ₂ ) ₂ CO—. In some embodiments, the spacer is γ- _{aminobutanoyl, ie, —NH(CH 2} ) ₃ CO—. In some embodiments, the spacer is β-asparagyl, ie, —NH(CHCO ₂ H)(CH ₂ )CO—. In some embodiments, the spacer is —NH(CH ₂ ) ₂ CO—. In some embodiments, the spacer is glycyl. In some embodiments, the spacer is β-alanyl.

In some embodiments, the spacer is —NHCH(CO ₂ H)(CH ₂ ) ₂ CO—[N(R ₃ )((CH ₂ ) ₂ O(CH ₂ ) ₂ O) _q (CH ₂ )CO—] _{is r} . In some embodiments, the spacer is —NH(CH ₂ ) ₃ CO——[N(R ₃ )((CH ₂ ) ₂ O(CH ₂ ) ₂ O) _q (CH ₂ )CO—] _r . In some embodiments, the spacer is —NHCH(CO ₂ H)(CH ₂ ) ₂ CO—NH((CH ₂ ) ₂ O(CH ₂ ) ₂ O) ₂ (CH ₂ )CO—. In some embodiments, the spacer is —NH(CH ₂ ) ₃ CO—NH((CH ₂ ) ₂ O(CH ₂ )2O) ₂ (CH ₂ )CO—. In some embodiments, the spacer is —NHCH(CO ₂ H)CH ₂ CO——[N(R ₃ )((CH ₂ ) ₂ O(CH ₂ ) ₂ O) _q (CH ₂ )CO—] _r . In some embodiments, the spacer is —NH(CH ₂ ) ₂ CO——[N(R ₃ )((CH ₂ ) ₂ O(CH ₂ ) ₂ O) _q (CH ₂ )CO—] _r .

In some embodiments, the spacer comprises a divalent group of Formula (XIII):

during the meal,

Y is selected from the group consisting of γGlu, Asp, Lys and Gly;

n is 1 to 10.

In some embodiments, Y is selected from the group consisting of γGlu and Gly. In some embodiments, Y is γGlu. In some embodiments, Y is Gly.

In some embodiments, the spacer forms a bridge between the amino group of the disclosed polypeptides and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with an amino group of the disclosed polypeptide and the other end of the spacer forms a covalent bond with a hydrogen atom or protecting group.

In some embodiments, the spacer comprises a divalent group of Formula (XIV):

during the meal,

n is 1 to 10.

In some embodiments, n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

In some embodiments, the spacer forms a bridge between the amino group of the disclosed polypeptides and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with an amino group of the disclosed polypeptide and the other end of the spacer forms a covalent bond with a hydrogen atom or protecting group.

In some embodiments, the spacer comprises a divalent group of Formula (XV):

during the meal,

n is 1 to 10.

In some embodiments, (γGlu) _n is γGlu; 2(γGlu); 3(γGlu); 4(γGlu) (SEQ ID NO: 313); and 5(γGlu) (SEQ ID NO: 314). In some embodiments, -(γGlu) _n -(Gly)-(“(γGlu) _n -(Gly)” set forth in SEQ ID NO: 312) is 2(γGlu),Gly; and 3 (γGlu), Gly (SEQ ID NO: 315).

In some embodiments, the spacer forms a bridge between the amino group of the disclosed polypeptides and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with an amino group of the disclosed polypeptide and the other end of the spacer forms a covalent bond with a hydrogen atom or protecting group.

In some embodiments, the spacer comprises a divalent group of Formula (XVI):

during the meal,

n is 1 to 10.

In some embodiments, the spacer forms a bridge between the amino group of the disclosed polypeptides and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with an amino group of the disclosed polypeptide and the other end of the spacer forms a covalent bond with a hydrogen atom or protecting group.

In some embodiments of Formula (XIII), Formula (XIV), Formula (XV), or Formula (XVI), n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

In some embodiments, the spacer comprises a divalent group of Formula (XVII):

during the meal,

Y1 is selected from the group consisting of γGlu, Asp and Gly;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

V is -[COCH ₂ (O(CH ₂ ) ₂ ) _t OCH ₂ NH]-, t is 1 to 8;

r is 1 to 8;

n1 is 0 to 10; And

n2 is 0 to 10.

In some embodiments, the spacer forms a bridge between the amino group of the disclosed polypeptides and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with an amino group of the disclosed polypeptide and the other end of the spacer forms a covalent bond with a hydrogen atom or protecting group.

In some embodiments, the spacer comprises a divalent group of Formula XVIII:

during the meal,

Y1 is selected from the group consisting of γGlu, Asp and Gly;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

dpeg is -[CO(CH ₂ )O(CH ₂ ) ₂ O(CH ₂ )NH]-;

r is 1 to 8;

n1 is 0 to 10; And

n2 is 0 to 10.

In some embodiments, -(Y1) _n1 -(dpeg) _r -(Y2) _n2 - is γGlu,dpeg,dpeg,γGlu; γGlu, dpeg, dpeg, 2 (γGlu); γGlu, dpeg, dpeg, 3 (γGlu); γGlu, dpeg, dpeg, 4 (γGlu); 2(γGlu), dpeg, dpeg, γGlu; and 2(γGlu),dpeg,γGlu.

In some embodiments, the spacer forms a bridge between the amino group of the disclosed polypeptides and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with an amino group of the disclosed polypeptide and the other end of the spacer forms a covalent bond with a hydrogen atom or protecting group.

Accordingly, in some embodiments, an isolated polypeptide provided herein comprises SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208 and SEQ ID NO: 209 comprising an amino acid sequence selected from the group consisting of an amino acid sequence represented by a consensus sequence selected from the group consisting of, or a pharmaceutically acceptable salt thereof, wherein the isolated peptide adds a lipophilic substituent and optionally a spacer.

In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent of Formula (I). In an embodiment, the isolated polypeptide further comprises a lipophilic substituent and a spacer selected from the group consisting of Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, and Formula X, and the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula III. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VI. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VII.

In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 200, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent of Formula (I). In an embodiment, the isolated polypeptide further comprises a lipophilic substituent and a spacer selected from the group consisting of Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, and Formula X, and the amino acid sequence of SEQ ID NO: 200, or a pharmaceutically acceptable salt thereof. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 200, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula III. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 200, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VI. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 200, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VII.

In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent of Formula (I). In an embodiment, the isolated polypeptide further comprises a lipophilic substituent and a spacer selected from the group consisting of Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, and Formula X, and the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula III. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VI. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VII.

In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 206, further comprising a lipophilic substituent of Formula (I), or a pharmaceutically acceptable salt thereof. In an embodiment, the isolated polypeptide further comprises a lipophilic substituent and a spacer selected from the group consisting of Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, and Formula X, and the amino acid sequence of SEQ ID NO: 206, or a pharmaceutically acceptable salt thereof. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 206, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula III. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 206, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VI. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 206, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VII.

In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent of Formula (I). In an embodiment, the isolated polypeptide further comprises a lipophilic substituent and a spacer selected from the group consisting of Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, and Formula X, and the amino acid sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula III. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VI. In an embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof, further comprising a spacer and a lipophilic substituent of Formula VII.

In some embodiments, an isolated polypeptide provided herein comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO:200, or a pharmaceutically acceptable salt thereof:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200 ), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; X ₈ is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₃ is A, S, E or K; X ₁₄ is N, n, d, Y or Q; X ₁₅ is E, F, f, Y, I, k, K or Aib; X ₁₆ is k, K, L, Aib, N -MeL or 1; X ₁₇ is H, V, Q, R, k, K or Aib; X ₁₈ is K, H or R; X ₁₉ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ is P, R or K; X ₃₁ is k, K or N; and X ₃₅ is e, E or N; each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge; provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K; The isolated peptide further comprises a lipophilic substituent of formula (I):

during the meal,

Z is —CH ₃ or —CO ₂ H; And

m is 4 to 24.

In some embodiments, an isolated polypeptide provided herein comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO:200, or a pharmaceutically acceptable salt thereof:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200 ), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; X ₈ is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₃ is A, S, E or K; X ₁₄ is N, n, d, Y or Q; X ₁₅ is E, F, f, Y, I, k, K or Aib; X ₁₆ is k, K, L, Aib, N -MeL or 1; X ₁₇ is H, V, Q, R, k, K or Aib; X ₁₈ is K, H or R; X ₁₉ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ is P, R or K; X ₃₁ is k, K or N; and X ₃₅ is e, E or N; each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge; provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K; The isolated peptide further comprises a lipophilic substituent of Formula III and a spacer:

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24; And

n is 1 to 10.

In some embodiments, an isolated polypeptide provided herein comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO:200, or a pharmaceutically acceptable salt thereof:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200 ), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; X ₈ is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₃ is A, S, E or K; X ₁₄ is N, n, d, Y or Q; X ₁₅ is E, F, f, Y, I, k, K or Aib; X ₁₆ is k, K, L, Aib, N -MeL or 1; X ₁₇ is H, V, Q, R, k, K or Aib; X ₁₈ is K, H or R; X ₁₉ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ is P, R or K; X ₃₁ is k, K or N; and X ₃₅ is e, E or N; each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge; provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K; The isolated peptide further comprises a lipophilic substituent of formula (VI) and a spacer:

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24;

Y1 is selected from the group consisting of γGlu, Asp and Gly;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

V is -[COCH ₂ (O(CH ₂ ) ₂ ) _t OCH ₂ NH]-, t is 1 to 8;

r is 1 to 8;

n1 is 0 to 10; And

n2 is 0 to 10.

In some embodiments, an isolated polypeptide provided herein comprises an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO:200, or a pharmaceutically acceptable salt thereof:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200 ), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; X ₈ is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₃ is A, S, E or K; X ₁₄ is N, n, d, Y or Q; X ₁₅ is E, F, f, Y, I, k, K or Aib; X ₁₆ is k, K, L, Aib, N -MeL or 1; X ₁₇ is H, V, Q, R, k, K or Aib; X ₁₈ is K, H or R; X ₁₉ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ is P, R or K; X ₃₁ is k, K or N; and X ₃₅ is e, E or N; each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge; provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K; The isolated peptide further comprises a lipophilic substituent of formula (VII) and a spacer:

during the meal,

Z is —CH ₃ or —CO ₂ H;

m is 4 to 24;

Y1 is selected from the group consisting of γGlu, Asp and Gly;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

dpeg is -[CO(CH ₂ )O(CH ₂ ) ₂ O(CH ₂ )NH]-;

r is 1 to 8;

n1 is 0 to 10; And

n2 is 0 to 10.

In some embodiments, the isolated polypeptide comprises a lipophilic substituent at _{position X 1} , X ₁₅ , or X _{16 .}

In some embodiments, _{the isolated polypeptide comprising a lipophilic substituent at position X 1} , X ₁₅ or X ₁₆ is conjugated via a lysine at that position.

In some embodiments, the isolated polypeptide comprises a lipophilic substituent at _{position X 1 .}

In some embodiments, the isolated polypeptide comprises a lipophilic substituent at _{position X 15 .}

In some embodiments, the isolated polypeptide comprises a lipophilic substituent at _{position X 16 .}

In some embodiments, _{the isolated polypeptide comprising a lipophilic substituent at position X 1} is conjugated via a lysine at that position.

In some embodiments, _{the isolated polypeptide comprising a lipophilic substituent at position X 15} is conjugated via a lysine at that position.

In some embodiments, _{the isolated polypeptide comprising a lipophilic substituent at position X 16} is conjugated via a lysine at that position.

In some embodiments, an isolated polypeptide of the present disclosure comprises the following amino acid sequence: SC*NTSTC*ATQRLANEk*((γGlu) ₂ -CO(CH ₂ ) ₁₄ CH _{3 , also referred to herein as Compound A27} ) HKSSNNFGPILPP TKVGSETY-NH ₂ (SEQ ID NO: 27).

In some embodiments, an isolated polypeptide of the present disclosure comprises the following amino acid sequence: K*((γGlu) ₂ (CO(CH ₂ ) ₁₈ CO ₂ H)), also referred to herein as Compound A64 C*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 64).

In some embodiments, an isolated polypeptide of the present disclosure comprises the amino acid sequence of: K*((γGlu) ₂ (CO(CH ₂ ) ₁₆ CO ₂ H)), also referred to herein as Compound Compound A65 C*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 65).

In some embodiments, an isolated polypeptide of the present disclosure comprises the following amino acid sequence: K*(γGlu-CO(CH ₂ ) ₁₆ CO ₂ H)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY, also referred to herein as Compound A109 -NH ₂ (SEQ ID NO: 109).

6. Exemplary Compounds: Amylin Analog Polypeptides

In some embodiments, an isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the following peptides listed in Table 3:

main:

각각의 K*는 독립적으로 선택적으로 스페이서를 통해 친유성 치환체에 선택적으로 공유결합된 L-라이신을 나타내며;

each K* independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

각각의 k*는 독립적으로 선택적으로 스페이서를 통해 친유성 치환체에 선택적으로 공유결합된 D-라이신을 나타내고;

each k* independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;

2 및 7번 위치에서 2개의 시스테인 잔기(C*)는 선택적으로 이황화 브리지에 의해 추가로 결합되며;

two cysteine residues (C*) at positions 2 and 7 are optionally further linked by a disulfide bridge;

본 명세서에 사용되는 바와 같은, (γGlu)₂와 2(γGlu)는 둘 다 -(γGlu)-(γGlu)-를 의미하고; (γGlu)₃과 3(γGlu)은 둘 다 -(γGlu)-(γGlu)-(γGlu)- 등을 의미하며; 그리고

As used herein, (γGlu) ₂ and 2(γGlu) both mean -(γGlu)-(γGlu)-; (γGlu) ₃ and 3 (γGlu) both mean -(γGlu)-(γGlu)-(γGlu)- and the like; And

여기서, 변수는 주어진 식에서 1회 초과로 존재하며, 해당 변수의 각 경우는 독립적으로 결정된다. 예를 들어, -(Y)₃-기에 대해, Y는 γGlu, Asp, Lys 또는 Gly일 수 있고, 각각의 Y는 4개의 아미노산 중 하나가 되도록 독립적으로 선택된다. 따라서, 비제한적 예에 의해, -(Y)₃-은 -(γGlu)-(γGlu)-(γGlu)-, -(γGlu)-(Asp)-(γGlu)-, -(Gly)-(Asp)-(γGlu)- 또는 -(Gly)-(γGlu)-(γGlu)-일 수 있다.

where the variable occurs more than once in a given equation, and each instance of that variable is independently determined. For example, for a group -(Y) ₃ -, Y can be γGlu, Asp, Lys or Gly, each Y being independently selected to be one of four amino acids. Thus, by way of non-limiting example, -(Y) ₃ - is -(γGlu)-(γGlu)-(γGlu)-, -(γGlu)-(Asp)-(γGlu)-, -(Gly)-(Asp )-(γGlu)- or -(Gly)-(γGlu)-(γGlu)-.

In some embodiments, the present invention provides a compound set forth in Table 3 above, or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutically acceptable salt is acetate. In some embodiments, the pharmaceutically acceptable salt is a trifluoroacetic acid (TFA) salt. In some embodiments, the pharmaceutically acceptable salt is a hydrochloric acid (HCl) salt. In one embodiment, the compound is A27. In some embodiments, the compound is the acetate salt of Compound A27. In some embodiments, the compound is a TFA salt of Compound A27. In some embodiments, the compound is the HCl salt of Compound A27. In one embodiment, the compound is A57. In some embodiments, the compound is the acetate salt of Compound A57. In some embodiments, the compound is the TFA salt of Compound A57. In some embodiments, the compound is the HCl salt of Compound A57. In one embodiment, the compound is A64. In some embodiments, the compound is the acetate salt of Compound A64. In some embodiments, the compound is a TFA salt of Compound A64. In some embodiments, the compound is the HCl salt of Compound A64.

7. Polypeptide intermediates

In certain embodiments, the present invention also relates to synthetic intermediates of isolated polypeptides that are amylin analogs. In some embodiments, a polypeptide intermediate of the present disclosure is an isolated polypeptide comprising an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 210:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 210 ), here:

X ₁ is S, K, k, H or I;

X ₃ is N or S;

X ₅ is S or A;

X ₆ is T or S;

X ₈ is A or K;

X ₁₀ is Q or S;

X ₁₂ is L or K;

X ₁₃ is A, S, E or K;

X ₁₄ is N, n, d, Y or Q;

X ₁₅ is E, F, f, Y, I, k, K or α-aminoisobutyric acid (Aib);

X ₁₆ is k, K, L, Aib, N -methyl leucine ( N -MeL) or 1;

X ₁₇ is H, V, Q, R, k, K or Aib;

X ₁₈ is K, H or R;

X ₁₉ is S or Aib;

X ₂₀ is S or Aib;

X ₂₂ is N or E;

X ₂₉ is P, R or K;

X ₃₁ is k, K or N; And

X ₃₅ is e, E or N;

each K independently represents L -lysine optionally covalently bound to a protecting group or a spacer optionally covalently bound to a protecting group;

each k independently represents a D -lysine optionally covalently bound to a protecting group or a spacer optionally covalently bound to a protecting group;

the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge;

provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, X ₃₁ is K and X ₃₅ is E.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 210 include:

In some embodiments, the carboxy terminus amino acid 37 is Y-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k. In some embodiments, X ₁ is H. In some embodiments, X ₁ is I.

In some embodiments, X ₃ is N. In some embodiments, X ₃ is S.

In some embodiments, X ₅ is S. In some embodiments, X ₅ is A.

In some embodiments, X ₆ is T. In some embodiments, X ₆ is S.

In some embodiments, X ₈ is A. In some embodiments, X ₈ is K.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₂ is L. In some embodiments, X ₁₂ is K.

In some embodiments, X ₁₃ is A. In some embodiments, X ₁₃ is S. In some embodiments, X ₁₃ is E. In some embodiments, X ₁₃ is K.

In some embodiments, X ₁₄ is N. In some embodiments, X ₁₄ is n. In some embodiments, X ₁₄ is d. In some embodiments, X ₁₄ is Y. In some embodiments, X ₁₄ is Q.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F. In some embodiments, X ₁₅ is f. In some embodiments, X ₁₅ is Y. In some embodiments, X ₁₅ is I. In some embodiments, X ₁₅ is K. In some embodiments, X ₁₅ is k. In some embodiments, X ₁₅ is Aib.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is l. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k. In some embodiments, X ₁₆ is Aib. In some embodiments, X ₁₆ is N -MeL.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q. In some embodiments, X ₁₇ is R. In some embodiments, X ₁₇ is K. In some embodiments, X ₁₇ is k. In some embodiments, X ₁₇ is Aib.

In some embodiments, X ₁₈ is K. In some embodiments, X ₁₈ is H. In some embodiments, X ₁₈ is R.

In some embodiments, X ₁₉ is S. In some embodiments, X ₁₉ is Aib.

In some embodiments, X ₂₀ is S. In some embodiments, X ₂₀ is Aib.

In some embodiments, X ₂₂ is N. In some embodiments, X ₂₂ is E.

In some embodiments, X ₂₉ is P. In some embodiments, X ₂₉ is R. In some embodiments, X ₂₉ is K.

In some embodiments, X ₃₁ is k. In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is e. In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 210 include:

In some embodiments, X ₁ is S and X ₅ is S. In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₈ is K. In some embodiments, X ₁ is S and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E.

In some embodiments, X ₁ is K and X ₅ is S. In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₈ is K. In some embodiments, X ₁ is K and X ₃₁ is K. In some embodiments, X ₁ is K and X ₃₅ is E.

In some embodiments, X ₁ is k and X ₅ is S. In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₈ is K. In some embodiments, X ₁ is k and X ₃₁ is K. In some embodiments, X ₁ is k and X ₃₅ is E.

In some embodiments, X ₅ is S and X ₁₅ is E. In some embodiments, X ₅ is S and X ₁₀ is Q. In some embodiments, X ₅ is S and X ₁₆ is L. In some embodiments, X ₅ is S and X ₁₆ is k. In some embodiments, X ₅ is S and X ₁₇ is H. In some embodiments, X ₅ is S and X ₁₈ is K. In some embodiments, X ₅ is S and X ₃₁ is K. In some embodiments, X ₅ is S and X ₃₅ is E.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₈ is K. In some embodiments, X ₁₀ is Q and X ₃₁ is K. In some embodiments, X ₁₀ is Q and X ₃₅ is E.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₈ is K. In some embodiments, X ₁₅ is E and X ₃₁ is K. In some embodiments, X ₁₅ is E and X ₃₅ is E.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₈ is K. In some embodiments, X ₁₆ is L and X ₃₁ is K. In some embodiments, X ₁₆ is L and X ₃₅ is E.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₈ is K. In some embodiments, X ₁₆ is k and X ₃₁ is K. In some embodiments, X ₁₆ is k and X ₃₅ is E.

In some embodiments, X ₁₇ is H and X ₁₈ is K. In some embodiments, X ₁₇ is H and X ₃₁ is K. In some embodiments, X ₁₇ is H and X ₃₅ is E.

In some embodiments, X ₁₈ is K and X ₃₁ is K. In some embodiments, X ₁₈ is K and X ₃₅ is E.

In some embodiments, X ₃₁ is K and X ₃₅ is E.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 210 include:

In some embodiments, X ₁ is S, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E.

In some embodiments, X ₁ is K, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₁ is k, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is k. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₇ is H. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₈ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₁ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₅ is E.

In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₁₀ is Q, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₀ is Q, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₀ is Q, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₅ is E.

In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₅ is E.

In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, the polypeptide intermediate of the present disclosure is an isolated polypeptide comprising an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 211:

X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 211), wherein:

X ₁ is S, K or k;

X ₁₀ is Q or S;

X ₁₅ is E or F;

X ₁₆ is L, K or k;

X ₁₇ is H, V or Q; And

X ₃₅ is E or N;

each K independently represents L -lysine optionally covalently bound to a protecting group or a spacer optionally covalently bound to a protecting group;

each K independently represents D -lysine optionally covalently bound to a protecting group or a spacer optionally covalently bound to a protecting group; And

The _{two cysteine residues of X 1} CNTSTC (SEQ ID NO: 309) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 211 include:

In some embodiments, the carboxy terminus amino acid 37 is Y-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 211 include:

In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₀ is S. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₅ is F. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is K. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₇ is V. In some embodiments, X ₁ is S and X ₁₇ is Q. In some embodiments, X ₁ is S and X ₃₅ is E. In some embodiments, X ₁ is S and X ₃₅ is N.

In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₀ is S. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₅ is F. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is K. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₇ is V. In some embodiments, X ₁ is K and X ₁₇ is Q. In some embodiments, X ₁ is K and X ₃₅ is E. In some embodiments, X ₁ is K and X ₃₅ is N.

In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₀ is S. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₅ is F. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is K. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₇ is V. In some embodiments, X ₁ is k and X ₁₇ is Q. In some embodiments, X ₁ is k and X ₃₅ is E. In some embodiments, X ₁ is k and X ₃₅ is N.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₅ is F. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is K. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₇ is V. In some embodiments, X ₁₀ is Q and X ₁₇ is Q. In some embodiments, X ₁₀ is Q and X ₃₅ is E. In some embodiments, X ₁₀ is Q and X ₃₅ is N.

In some embodiments, X ₁₀ is S and X ₁₅ is E. In some embodiments, X ₁₀ is S and X ₁₅ is F. In some embodiments, X ₁₀ is S and X ₁₆ is L. In some embodiments, X ₁₀ is S and X ₁₆ is K. In some embodiments, X ₁₀ is S and X ₁₆ is k. In some embodiments, X ₁₀ is S and X ₁₇ is H. In some embodiments, X ₁₀ is S and X ₁₇ is V. In some embodiments, X ₁₀ is S and X ₁₇ is Q. In some embodiments, X ₁₀ is S and X ₃₅ is E. In some embodiments, X ₁₀ is S and X ₃₅ is N.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₅ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₇ is V. In some embodiments, X ₁₆ is k and X ₁₇ is Q. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, a polypeptide intermediate of the present disclosure is an isolated polypeptide comprising an amino acid sequence selected from the group consisting of the amino acid sequence represented by the consensus sequence of SEQ ID NO: 212:

SCNTSTCATQRLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 212), wherein:

X ₁₅ is E or F;

X ₁₆ is L, K or k;

X ₁₇ is H, V or Q; And

X ₃₅ is E or N;

each K independently represents L -lysine optionally covalently bound to a protecting group or a spacer optionally covalently bound to a protecting group;

each K independently represents D -lysine optionally covalently bound to a protecting group or a spacer optionally covalently bound to a protecting group; And

The two cysteine residues of SCNTSTC (SEQ ID NO: 310) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 212 include:

In some embodiments, the carboxy terminus amino acid 37 is Y-(NH ₂ ). In some embodiments, the carboxy terminus amino acid 37 is Y-(OH).

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₅ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₇ is V. In some embodiments, X ₁₆ is k and X ₁₇ is Q. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, the isolated peptide comprising the amino acid sequence of SEQ ID NO: 210, SEQ ID NO: 211, or SEQ ID NO: 212 further comprises a protecting group or spacer optionally linked to a protecting group. In some embodiments, the isolated peptide further comprises a protecting group. In some embodiments, the isolated peptide further comprises a spacer. In some embodiments, the isolated peptide further comprises a spacer bound to a protecting group.

In some embodiments, the isolated polypeptide comprises a protecting group or a spacer optionally linked to a protecting group at _{position X 1} , X ₁₅ or X _{16 .}

In some embodiments, the isolated polypeptide comprising a protecting group or a spacer optionally bound to a protecting group at _{positions X 1} , X _{15 ,} or X _{16 is conjugated via a lysine at that position.}

In some embodiments, the isolated polypeptide comprises a protecting group or a spacer optionally linked to a protecting group at _{position X 1 .}

In some embodiments, the isolated polypeptide comprises a spacer optionally linked to a protecting group or a protecting group at _{position X 15 .}

In some embodiments, the isolated polypeptide comprises a spacer optionally linked to a protecting group or a protecting group at _{position X 16 .}

In some embodiments, an isolated polypeptide comprising a protecting group or a spacer optionally bound to a protecting group at _{position X 1 is conjugated via a lysine at that position.}

In some embodiments, the isolated polypeptide comprising a protecting group or a spacer optionally bound to a protecting group at _{position X 15 is conjugated via a lysine at that position.}

In some embodiments, the isolated polypeptide comprising a protecting group or a spacer optionally bound to a protecting group at _{position X 16 is conjugated via a lysine at that position.}

In some embodiments, the protecting group is acetyl, allyloxycarbonyl, benzyl, Boc, Cbz, Dmb, (dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl, Fmoc, tert-butyl, or tri is selected from the group consisting of tyl. In some embodiments, the protecting group is acetyl, allyloxycarbonyl, dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl, Fmoc, tert-butyl, or trityl. In some embodiments, the protecting group is acetyl, allyloxycarbonyl, dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl or Fmoc.

In some embodiments, the spacer is selected from the group consisting of Formula XI, Formula XII, Formula XIII, Formula XIV, Formula XV, Formula XVI, Formula XVII, and Formula XVIII. In some embodiments, the isolated polypeptide further comprises a spacer of Formula (XIII). In some embodiments, the isolated polypeptide further comprises a spacer of Formula (XIV). In some embodiments, the isolated polypeptide further comprises a spacer of Formula (XVII). In some embodiments, the isolated polypeptide further comprises a spacer of Formula (XVIII). In some embodiments, the spacer is bonded to a protecting group. In some embodiments, the protecting group is acetyl, allyloxycarbonyl, benzyl, Boc, Cbz, Dmb, (dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl, Fmoc, tert-butyl, or tri is selected from the group consisting of tyl. In some embodiments, the protecting group is acetyl, allyloxycarbonyl, dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl, Fmoc, tert-butyl, or trityl. In some embodiments, the protecting group is acetyl, trityl or tert-butyl. In some embodiments, the spacer is not bound to a protecting group.

Exemplary polypeptide intermediates

In some embodiments, an isolated polypeptide of the present disclosure comprises an amino acid sequence selected from the group consisting of the following peptides listed in Table 4:

In some embodiments, the invention provides the peptide intermediates set forth in Table 4 above. In some embodiments, the polypeptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 129, SEQ ID NO: 144, SEQ ID NO: 145, or SEQ ID NO: 156. In some embodiments, the polypeptide intermediate has the amino acid sequence of SEQ ID NO: 129. In some embodiments, the polypeptide intermediate has the amino acid sequence of SEQ ID NO:145. In some embodiments, the polypeptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 129, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, or SEQ ID NO: 150. In some embodiments, the polypeptide intermediate has the amino acid sequence of SEQ ID NO: 147. In some embodiments, the polypeptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, or SEQ ID NO: 154. In some embodiments, the polypeptide intermediate has the amino acid sequence of SEQ ID NO: 151.

In some embodiments, the polypeptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 155 or SEQ ID NO: 156. In some embodiments, the polypeptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 155. In some embodiments, the poly*tide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, or SEQ ID NO: 160. In some embodiments, the polypeptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 157. In some embodiments, the polypeptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, or SEQ ID NO: 164. In some embodiments, the polypeptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 164.

8. Uses, Formulations and Administration

How to use

According to another embodiment, the present invention provides a method for treating a metabolic disease or disorder in a subject in need thereof comprising providing to the subject an effective amount of an amylin analog polypeptide of the present disclosure or a pharmaceutical composition thereof. It's about how to treat. Metabolic diseases or disorders include type 1 diabetes, type 2 diabetes, and obesity. Additionally, the present invention relates to a method of achieving weight loss in a subject, including a diabetic subject, comprising providing to the subject an effective amount of an amylin analog polypeptide of the present disclosure.

Amylin analog polypeptides of the present disclosure are particularly useful for treating diabetes, the method comprising providing an effective amount of an amylin analog polypeptide to a diabetic subject. In some embodiments, an amylin analog polypeptide of the present disclosure is used for the treatment of a subject having type 1 or type 2 diabetes to control or reduce blood glucose levels in the subject, wherein the blood glucose level is glycated hemoglobin (hemoglobin). It can be monitored or approximated based on the measured blood concentration of A1c,　HbA1c).

(i) In some embodiments, an amylin analog polypeptide of the present disclosure is used for the treatment of a subject having type 1 diabetes;

(ii) In some embodiments, an amylin analog polypeptide of the present disclosure is used for the treatment of a subject having type 2 diabetes;

(iii) In some embodiments, an amylin analog polypeptide of the present disclosure is used for the treatment of obesity; And

(iv) In some embodiments, an amylin analog polypeptide of the present disclosure is used to provide weight loss in a subject, such as a diabetic subject,

The amylin analog polypeptide of use (i), (ii), (iii) or (iv) is SEQ ID NO: 199, 200, 204, 206, 127, 57, 128, 129, 43, 209, 130, 64, 65 , 131, 109 and 27;

In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having type 1 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:27. In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having type 1 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:64. In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having type 1 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:65. In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having type 1 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:131.

In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having type 2 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:27. In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having type 2 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:64. In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having type 2 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:65. In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having type 2 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:131.

In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having obesity, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:27. In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having obesity, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:64. In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having obesity, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:65. In some embodiments, the amylin analog polypeptide is used for the treatment of a subject having obesity, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:131.

In some embodiments, the amylin analog polypeptide is used to provide weight loss for a subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:27. In some embodiments, the amylin analog polypeptide is used to provide weight loss for a subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:64. In some embodiments, the amylin analog polypeptide is used to provide weight loss for a subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:65. In some embodiments, the amylin analog polypeptide is used to provide weight loss for a subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO:131.

Amylin analog polypeptides of the present disclosure, such as insulin, are provided (ie, administered to) a diabetic subject to maintain, control, or reduce blood glucose levels in the subject. Diabetic subjects treated with an amylin analog polypeptide of the present disclosure as an adjunct to insulin therapy are at risk of hypoglycemia (ie, low blood sugar), particularly severe hypoglycemia. Thus, reducing the dietary insulin dose for diabetic subjects upon treatment with an amylin analog polypeptide of the present disclosure is intended to reduce the risk of hypoglycemia, particularly severe hypoglycemia.

Severe hypoglycemia, as used herein, is an episode of hypoglycemia that requires the assistance of another individual (including helping to administer oral carbohydrates) or that requires administration of glucagon, intravenous glucose, or other medical intervention. refers to

Accordingly, administration of an amylin analog polypeptide of the present disclosure as an adjunct to insulin therapy, particularly dietary insulin therapy, generally requires a reduction in the dose of dietary insulin necessary to adequately maintain healthy blood sugar in a subject. In other words, type 1 or type 2 diabetes, which is already self-administered with dietary insulin at a certain dose prior to initiation of treatment with the amylin analog polypeptide of the present disclosure, when starting treatment with the amylin analogue polypeptide of the present disclosure They will reduce the dose of dietary insulin (eg by up to 25%, 50%, 75% or 100%) as they continue self-administration.

In some embodiments, the method comprises providing an amylin analog polypeptide of the disclosure or a pharmaceutical composition thereof via injection to a subject in need thereof. In some embodiments, the method comprises providing an amylin analog polypeptide of the present disclosure or a pharmaceutical composition thereof formulated for oral administration to a subject in need thereof.

In some embodiments, the method comprises providing an amylin analog polypeptide of the disclosure or a pharmaceutical composition thereof via transplantation to a subject in need thereof. In some embodiments, the method comprises providing continuous delivery of an amylin analog polypeptide from an osmotic delivery device to a subject in need thereof. A delivery device, such as a canine osmotic delivery device, comprises sufficient amylin analog polypeptides of the present disclosure for continuous administration for up to 3 months, 6 months, 9 months, 12 months, 18 months or 24 months. In this way, continuous administration of an amylin analog polypeptide of the present disclosure via an osmotic delivery device eliminates daily or multiple daily dosing of an amylin analog polypeptide, such as pramlintide, present. For diabetes treated with pramlintide, the dosing of pramlintide before a meal should be adjusted according to meal insulin administered after a meal. In contrast, diabetes treated with an amylin analog polypeptide of the present disclosure via an osmotic delivery device receives continuous delivery of the amylin analog polypeptide and requires only reduced doses of dietary insulin to be administered.

Substantial steady state delivery of the amylin analog polypeptide from the osmotic delivery device is continuous over the administration period. In some embodiments, the subject or patient is a human subject or human patient.

In some embodiments of the invention, the administration period is, for example, at least about 3 months, at least about 3 months to about 1 year, at least about 4 months to about 1 year, at least about 5 months to about 1 year, at least about 6 months to about 1 year, at least about 8 months to about 1 year, at least about 9 months to about 1 year, at least about 10 months to about 1 year, at least about 1 year to about 2 years, at least about 2 years to about 3 is the year

In a further embodiment, the method of treatment of the present invention is achieved within about 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day or less after implantation of the osmotic delivery device in the subject (implantation of the osmotic delivery device) and provide a significant decrease in the fasting blood glucose concentration of the subject following implantation of the osmotic delivery device in the subject (relative to the subject's fasting blood glucose concentration before). A significant decrease in fasting blood glucose is typically either statistically significant as evidenced by application of an appropriate statistical test or is considered by a healthcare practitioner to be significant to a subject. A significant reduction in fasting plasma relative to pre-transplant baseline is typically maintained over the dosing period.

In some embodiments, the present invention relates to a method of treating a disease or condition in a subject in need thereof. The method comprises providing continuous delivery of the drug from the osmotic delivery device, wherein substantially steady state delivery of the drug at the therapeutic concentration is achieved in the subject. The substantially steady state of the drug from the osmotic delivery device is continuous over an administration period of at least about 3 months. A drug has a known or determined half-life in a typical subject. Humans are preferred subjects for the practice of the present invention. The present invention encompasses osmotic delivery devices comprising a drug effective for the treatment of a disease or condition as well as a drug for use in the methods of treating a disease or condition in a subject in need thereof. Advantages of the present invention include alleviation of peak-related drug toxicity and attenuation of trough-related suboptimal drug therapy.

In some embodiments, substantially steady state delivery of the drug at the therapeutic concentration is achieved within a period of about 1 month, 7 days, 5 days, 3 days, or 1 day after implantation of the osmotic delivery device in the subject.

The invention also provides a method of promoting weight loss in a subject in need thereof, a method of treating excess weight or obesity in a subject in need thereof, and/or a method of suppressing appetite in a subject in need thereof. provides The method comprises providing for delivery of an isolated amylin analog polypeptide. In some embodiments, the isolated amylin analog polypeptide is delivered continuously from an implantable osmotic delivery device. In some embodiments, substantially steady state delivery of the amylin analog polypeptide from the osmotic delivery device is achieved and is substantially continuous over the period of administration. In some embodiments, the subject is a human.

The present invention includes an osmotic delivery device comprising an isolated amylin analog polypeptide as well as an isolated amylin analog polypeptide for use in the methods in a subject in need thereof. The subject may have type 2 diabetes. A subject in need thereof, ie, a high baseline (HBL) subject, may have a baseline HbA1c% greater than 10.0%. The subject may not have already received a drug to treat type 2 diabetes mellitus.

In a further embodiment, the method of treatment of the present invention is administered within about 7 days or less after implantation of the osmotic delivery device in the subject, within about 6 days or less after implantation of the osmotic delivery device in the subject, or within about 5 days after implantation of the osmotic delivery device in the subject. within, within about 4 days or less after implantation of an osmotic delivery device in a subject, within about 3 days or less after implantation of an osmotic delivery device in a subject, within about 2 days or less after implantation of an osmotic delivery device in a subject, or osmotic delivery in a subject provides a significant reduction in the subject's fasting blood glucose following implantation of the osmotic delivery device in the subject (relative to the subject's fasting blood glucose concentration prior to implantation of the osmotic delivery device) achieved within about 1 day or less after implantation of the device. In a preferred embodiment of the present invention, a significant decrease in the fasting blood glucose concentration of the subject after implantation of the osmotic delivery device compared to the subject's fasting blood glucose concentration prior to implantation is within about 2 days or less, preferably after implantation of the osmotic delivery device in the subject by about within 1 day or less, or more preferably within about 1 day after implantation of the osmotic delivery device in the subject. A significant decrease in fasting blood glucose is typically either statistically significant as evidenced by application of an appropriate statistical test or is considered by a healthcare practitioner to be significant to a subject. A significant reduction in fasting plasma relative to pre-transplant baseline is typically maintained over the dosing period.

In embodiments of all aspects of the invention directed to a method of treating a disease or condition in a subject, the exemplary osmotic delivery device comprises: an impermeable reservoir comprising inner and outer surfaces and first and second open ends; a semipermeable membrane in sealing relationship with the first open end of the reservoir; an osmotic engine in the reservoir and an adjacent semipermeable membrane; a piston adjacent the osmotic engine, the piston forming a seal movable by an inner surface of the reservoir, the piston dividing the reservoir into a first chamber and a second chamber, the first chamber comprising the osmotic engine; a drug formulation or suspension formulation comprising a drug, wherein the second chamber contains the drug formulation or suspension formulation, the drug formulation or suspension formulation being flowable; and a diffusion regulator inserted into the second open end of the reservoir, the diffusion regulator adjacent the suspension formulation. In a preferred embodiment, the reservoir comprises titanium or a titanium alloy.

In embodiments of all aspects of the invention directed to a method of treating a disease or condition in a subject, the drug formulation may comprise a drug and vehicle formulation. Alternatively, suspension formulations are used in the present methods and may include particle formulations including, for example, drug and vehicle formulations. Vehicle formulations for use in forming the suspension formulations of the present invention may include, for example, a solvent and a polymer.

The reservoir of the osmotic delivery device may comprise, for example, titanium or a titanium alloy.

In embodiments of all aspects of the invention, the implanted osmotic delivery device may be used to provide subcutaneous delivery.

In embodiments of all aspects of the invention, the continuous delivery may be, for example, a zero-order controlled continuous delivery.

combination

In some embodiments, an amylin analog polypeptide of the present disclosure is co-formulated in combination with a second agent. In some embodiments, an amylin analog polypeptide of the disclosure is co-formulated in combination with a second agent, wherein the second agent is an insulinotropic peptide. In some embodiments, an amylin analog polypeptide of the disclosure is coformulated in combination with a second agent, wherein the second agent is a GLP-1 receptor agonist. In some embodiments, an amylin analog polypeptide of the present disclosure is coformulated in combination with a second agent, wherein the second agent is a (GLP-1) agonist, such as exenatide, a derivative of exenatide, exenatide It is an analogue of cenataide or semaglutide. In some embodiments, the GLP-1 receptor agonist is exenatide. In some embodiments, the GLP-1 receptor agonist is semaglutide.

In some embodiments, an amylin analog polypeptide of the present disclosure is administered to a subject in combination with a second agent without being co-formulated with the second agent, wherein the second agent is a (GLP-1) agonist, such as , exenatide, a derivative of exenatide, an analog of exenatide, or semaglutide.

In some embodiments, an amylin analog polypeptide of the disclosure is co-administered in combination with insulin or an insulin derivative. In some embodiments, an amylin analog polypeptide of the present disclosure is co-formulated in combination with a long acting basal insulin or long acting basal insulin derivative.

In some embodiments, an amylin analog polypeptide of the present disclosure is administered to a subject in combination with insulin or an insulin derivative, ie, as an adjunct to insulin therapy, without being co-formulated with the insulin or insulin derivative. In some embodiments, an amylin analog peptide of the present disclosure is administered to a subject in combination with a meal insulin, without being co-formulated with the insulin or insulin derivative. In some embodiments, the subject has type 1 diabetes. In some embodiments, the subject has type 2 diabetes.

In some embodiments, an amylin analog polypeptide of the present disclosure is co-administered with insulin or an insulin derivative to a human patient to provide a so-called dual-hormonal “artificial pancreas” therapy. In some embodiments, an amylin analog polypeptide of the present disclosure is co-administered to a subject in combination with an insulin or insulin derivative to provide a dual hormone “artificial pancreas” therapy without being co-formulated with the insulin or insulin derivative. do. In some embodiments, the amylin analog polypeptides of the present disclosure are co-formulated with insulin or an insulin derivative, and thus administered one by one to the subject in combination with the insulin or insulin derivative to provide a dual hormone "artificial pancreas" therapy. In some embodiments, the artificial pancreas therapy comprises a fast-acting insulin or a fast-acting insulin derivative. In some embodiments, the artificial pancreas therapy comprises a long-acting or basal insulin or a long-acting or basal insulin derivative.

In some embodiments, any amylin analog of the present disclosure is formulated in combination with exenatide. In some embodiments, any amylin analog of the present disclosure is formulated in combination with a GLP-1 receptor agonist.

Some embodiments of the invention provide a second therapeutic agent, such as a second polypeptide, such as, but not limited to, insulinotropic peptides, peptide hormones such as glucagon and incretin mimetics (eg, GLP- 1 receptor agonists such as exenatide) as well as peptide analogs and peptide derivatives thereof; PYY (also known as peptide YY, peptide tyrosine tyrosine), as well as peptide analogs and peptide derivatives thereof, such as PYY(3-36); oxyntomodulin as well as its peptide analogs and peptide derivatives); and the use of amylin analog polypeptides in combination with gastric inhibitory peptides (GIPs) as well as peptide analogs and peptide derivatives thereof. In some embodiments, a pharmaceutical composition comprising an amylin analog polypeptide in combination with a GLP-1 receptor agonist is used to treat type 2 diabetes.

GLP-1, which contains three types of peptides, GLP-1 (1-37), GLP-1 (7-37) and GLP-1 (7-36) amide, as well as a peptide analog of GLP-1, secretes insulin. has been shown to stimulate (i.e., insulinotropic), which induces glucose uptake by cells and results in a decrease in serum glucose concentration (see, e.g., Mojsov, S., Int. J. Peptide Protein Research, 40:333-343 (1992)).

Numerous GLP-1 receptor agonists (eg, GLP-1 peptide derivatives and peptide analogs) that demonstrate insulinotropic action are known in the art (eg, US Pat. Nos. 5,118,666; 5,120,712; 5,512,549; 5,545,618; 5,574,008; 5,574,008; 5,614,492; 5,958,909; 6,191,102; 6,268,343; 6,329,336; 6,451,974; 6,706,689; 6,720,407; 6,821,949; 6,849,708; 6,849,714; 6,887,470; 6,887,849; 6,903,186; 7,022,674; 7,041,646; 7,084,243; 7,138,486; 7,141,547; 7,144,863; and 7,199,217) as well as clinical trials (eg, taspoglutide and albiglutide). An example of a GLP-1 receptor agonist in the practice of the present invention is Victoza® (Novo Nordisk A/S, Bagsverd, Denmark) (liraglutide; US Pat. Nos. 6,268,343, 6,458,924 and 7,235,627). Once daily injectable Victoza® (liraglutide) is commercially available in the United States, Europe and Japan. Another example of a GLP-1 receptor agonist is Ozempic® (Novo Nordisk A/S, Bagsverd, Denmark) (semaglutide). For ease of reference herein, the family of GLP-1 receptor agonists with insulinotropic activity, GLP-1 peptides, GLP-1 peptide derivatives and GLP-1 peptide analogs are collectively referred to as “GLP-1”. .

The molecule exenatide has the amino acid sequence of exendin-4 (Kolterman OG, 　 et al., J. Clin. Endocrinol. Metab. 88(7):3082-9 (2003)), and is not suitable for chemical synthesis or recombinant expression. is created by For ease of reference herein, the family of exenatide peptides (including, for example, exendin-3, exendin-4 and exendin-4-amide), exenatide peptide derivatives and exenatide peptides Analogs are collectively referred to as “exenatides”.

Peptide YY (PYY) is a 36 amino acid residue peptide amide. PYY inhibits intestinal motility and blood flow (Laburthe, M., Trends Endocrinol Metab. 1(3):168-74 (1990)) and mediates intestinal secretion (Cox, HM, et al., Br J Pharmacol 101 ( 2):247-52 (1990); Playford, RJ, et al., Lancet 335(8705):1555-7 (1990)), stimulates net absorption (MacFayden, RJ, et al., Neuropeptides 7(3) ):219-27 (1986)) Two major in vivo variants, PYY(1-36) and PYY(3-36) have been identified (see, e.g., Eberlein, GA, et al., Peptides 10 (4), 797-803 (1989)]) The sequences of PYY as well as its peptide analogs and peptide derivatives are known in the art (eg, US Pat. Nos. 5,574,010 and 5,552,520).

Oxyntomodulin is a naturally occurring 37 amino acid peptide found in the colon that has been found to suppress appetite and facilitate weight loss (Wynne K, et al., Int J Obes (Lond) 30(12):1729-36). (2006)). The sequences of oxyntomodulin as well as its peptide analogs and peptide derivatives are known in the art (eg, Bataille D, et al., Peptides 2(Suppl 2):41-44 (1981)); and US Patent Publication Nos. 2005/0070469 and 2006/0094652).

Gastric inhibitory peptides (GIPs) are insulinotropic peptide hormones (Efendic, S., et al., Horm Metab Res. 36:742-6 (2004)), and the mucous membranes of the duodenum and jejunum in response to absorbed fats and carbohydrates. secreted by the pancreas to stimulate the pancreas to secrete insulin. GIP is circulated as a biologically active 42-amino acid peptide. GIP is also known as a glucose-dependent insulinotropic protein. GIP is a 42-amino acid gastrointestinal regulatory peptide that stimulates insulin secretion from pancreatic beta cells in the presence of glucose (Tseng, C., et al., PNAS 90:1992-1996 (1993)). The sequences of GIP as well as its peptide analogs and peptide derivatives are known in the art (see, e.g., Meier JJ, Diabetes Metab Res Rev. 21(2):91-117 (2005) and Efendic S., Horm). Metab Res. 36(11-12):742-6 (2004)]).

Glucagon is a peptide hormone produced by the alpha cells of the pancreas, which raises the concentration of glucose in the bloodstream. Its effect is the opposite of insulin, which lowers glucose levels. The pancreas releases glucagon when the concentration of glucose in the bloodstream drops too low. Glucagon causes the liver to convert stored glycogen into glucose, which is released into the bloodstream. High blood sugar levels stimulate insulin release. Insulin causes glucose to be consumed and used by insulin-dependent tissues. Thus, glucagon and insulin are part of a feedback system that maintains blood sugar levels at stable levels.

Pharmaceutically Acceptable Compositions

According to another embodiment, the present invention provides a composition comprising a compound of the present invention, ie, an isolated polypeptide, or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant or vehicle. The amount of the compound in the composition of the present invention is such that it is effective to measurably activate one or more amylin and/or calcitonin receptors in a biological sample or in a patient. In certain embodiments, the amount of the compound in a composition of the invention is such that it measurably activates human amylin 3 receptor (hAMY3) and/or human calcitonin receptor (hCTR) in the absence or presence of human serum albumin in a biological sample or in a patient. is effective for In certain embodiments, the compositions of the present invention are formulated for administration to a patient in need thereof. In some embodiments, the compositions of the present invention are formulated for injectable administration to a patient. In some embodiments, the compositions of the present invention are formulated for administration to a patient via an implantable delivery device, such as an osmotic delivery device.

The term “patient” or “subject” as used herein refers to an animal, preferably a mammal, most preferably a human.

"Pharmaceutically acceptable derivative" refers to any non-toxic salt of a compound of the present invention, which upon administration to a recipient, directly or indirectly, is capable of providing a compound of the present invention or an inhibitory active metabolite or residue thereof, esters, salts of esters or other derivatives.

The isolated polypeptides of the present disclosure (also referred to herein as "active compounds") and derivatives, fragments, analogs and homologs thereof can be incorporated into pharmaceutical compositions suitable for administration. Such compositions typically include an isolated polypeptide, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like suitable for pharmaceutical administration. It is intended Suitable carriers are described in the latest edition of Remington's Pharmaceutical Sciences, a standard reference in the art, incorporated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional medium or agent is inappropriate for the active compound, its use in the compositions is contemplated. Supplementary active compounds may also be incorporated into the compositions.

The pharmaceutical composition of the present invention is formulated to be suitable for its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subdermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e. topical), transmucosal, rectal, or combinations thereof. do. In some embodiments, the pharmaceutical composition or isolated polypeptide of the present disclosure is formulated for administration by topical administration. In some embodiments, the pharmaceutical composition or isolated polypeptide of the present disclosure is formulated for administration by inhalation administration. In some embodiments, the pharmaceutical composition is formulated for administration by a device suitable for subdermal or subcutaneous implantation and delivering the pharmaceutical composition subcutaneously or other suitable delivery mechanism. In some embodiments, the pharmaceutical composition is formulated for administration by an implantation device suitable for subdermal or subcutaneous implantation and delivering the pharmaceutical composition subcutaneously. In some embodiments, the pharmaceutical composition is formulated for administration by an osmotic delivery device that is suitable for subdermal or subcutaneous placement or other implantation and delivers the pharmaceutical composition subcutaneously, eg, an implantable osmotic delivery device. Solutions or suspensions used for parenteral application, intradermal application, subdermal application, subcutaneous application, or combinations thereof may contain the following components: a sterile diluent such as water for injection, saline solution, fixed oil, polyethylene glycol, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates and agents for the adjustment of isotonicity, such as sodium chloride or dextrose. The pH can be adjusted with acids or bases such as hydrochloric acid or sodium hydroxide. Parenteral preparations may be sealed in glass or plastic ampoules, episodic injections, or multi-dose vials.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (if water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL (BASF, Farsippany, NJ) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and must be fluid to the extent that easy syringeability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, a polyol (eg, glycerol, propylene glycol, and liquid polyethylene glycol, etc.), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions may be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, if necessary, followed by sterile filtration, if necessary. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation are vacuum drying and freeze drying obtained by adding the active ingredient powder to any additional desired ingredient from its previously sterile filtered solution.

Oral compositions generally include an inert diluent or edible carrier. They may be sealed in gelatin capsules or compressed into tablets. For oral therapeutic administration purposes, the active compounds may be incorporated with excipients and used in the form of tablets, buccal tablets or capsules. Oral compositions may also be prepared using a fluid carrier for use as a mouth wash, wherein the compound in the fluid carrier is applied orally, gargled, and spit or swallowed. Pharmaceutically suitable binder and/or adjuvant substances may be included as part of the composition. Tablets, pills, capsules, oral tablets and the like may contain any of the following ingredients, or compounds of similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch or lactose, disintegrants such as alginic acid, Primogel or corn starch; lubricants such as magnesium stearate or Sterotes; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylate or orange flavoring.

For administration by inhalation, the compounds are delivered in the form of an aerosol spray from a pressurized container or dispenser containing a suitable propellant, for example, a gas such as carbon dioxide, or a nebulizer.

Systemic administration may also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be penetrated are used in the formulation. Such penetrants are generally known in the art and include, for example, for transmucosal administration, detergents, bile salts and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels or creams as generally known in the art.

In one embodiment, the active compound is prepared with a carrier that will protect the compound against rapid clearance from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyacid anhydride, polyglycolic acid, collagen, polyorthoesters and polylactic acid. Methods for preparing such formulations will be apparent to those skilled in the art. The material is also commercially available from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions may also be used as pharmaceutically acceptable carriers. They can be prepared according to methods known to those skilled in the art, for example as described in US Pat. No. 4,522,811.

It is particularly advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suitable as single dosages for the subject being treated; Each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The details of the dosage unit form of the present invention are indicated by, and are directly dependent on, the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of formulating such active compounds for the treatment of individuals. .

The pharmaceutical composition may be included in a container, pack, or dispenser with instructions for administration.

drug particle formulation

In some embodiments, pharmaceutical compositions comprising any of the disclosed polypeptides formulated as trifluoroacetate, acetate or hydrochloride are provided. In some embodiments, pharmaceutical compositions comprising any of the disclosed polypeptides formulated as trifluoroacetate salts are provided. In some embodiments, pharmaceutical compositions comprising any of the disclosed polypeptides formulated as acetate salts are provided. In some embodiments, a pharmaceutical composition comprising any of the disclosed polypeptides formulated as a hydrochloride salt is provided.

A compound for use in the practice of the present invention, i.e., an isolated polypeptide or a pharmaceutically acceptable salt thereof, is typically added to a particle formulation, which is uniformly suspended, dissolved or dispersed in a suspension vehicle- It is used to produce the containing particles to form a suspension formulation. In some embodiments, the amylin analog polypeptide is formulated in a particle formulation and converted into particles (eg, spray dried). In some embodiments, the particles comprising the amylin analog polypeptide are suspended in a vehicle formulation to produce a suspension formulation of the vehicle comprising the amylin analog polypeptide and the suspended particles.

Preferably, the particle formulation is prepared by spray drying, lyophilization, desiccation, freeze drying, milling, granulation, ultrasonic droplet generation, crystallization, precipitation or other techniques available in the art for forming particles from a mixture of ingredients. It can be formed into particles using the same process as In one embodiment of the invention, the particles are spray dried. The particles are preferably substantially uniform in shape and size.

In some embodiments, the present invention provides drug particle formulations for pharmaceutical use. Particle formulations typically include a drug and one or more stabilizing ingredients (also referred to herein as “excipients”). Examples of stabilizing ingredients include, but are not limited to, carbohydrates, antioxidants, amino acids, buffers, inorganic compounds, and surfactants. The amount of stabilizer in a particle formulation can be determined empirically based on the activity of the stabilizer and the desired characteristics of the formulation in light of the teachings herein.

In certain embodiments, the particle formulation comprises from about 50% to about 90% drug, from about 50% to about 85% drug, from about 55% to about 90% drug, from about 60% to about 90% drug by weight. % drug, about 65% to about 85% drug, about 65% to about 90% drug, about 70% to about 90% drug, about 70% to about 85% drug, about 70% drug % to about 80% by weight drug or from about 70% to about 75% by weight drug.

Typically, the amount of carbohydrate in the particle formulation is determined by aggregation concerns. In general, the amount of carbohydrates should not be too high to avoid promoting crystal growth in the presence of water due to excess carbohydrates not bound to the drug.

Typically, the amount of antioxidant in the particle formulation is determined by oxidation concerns, while the amount of amino acids in the formulation is determined by oxidation concerns and/or the formability of the particles during spray drying.

Typically, the amount of buffer in the particle formulation is determined by pre-processing concerns, stability concerns, and the formability of the particles during spray drying. Buffers may be needed to stabilize the drug when all stabilizers are solubilized, for example during solution preparation and spray drying.

Examples of carbohydrates that may be included in the particle formulation include monosaccharides (eg fructose, maltose, galactose, glucose, D-mannose and sorbose), disaccharides (eg lactose, sucrose, trehalose and cellobiose). , polysaccharides (e.g., raffinose, melechtose, maltodextrin, dextrin and starch), and alditols (acyclic polyols; e.g., mannitol, xylitol, maltitol, lactitol, xylitol, sorbitol, pyranosyl sorbitol and myo oinositol), but are not limited thereto. Suitable carbohydrates include disaccharides and/or non-reducing sugars such as sucrose, trehalose and raffinose.

Examples of antioxidants that may be included in the particle formulation are methionine, ascorbic acid, sodium thiosulfate, catalase, platinum, ethylenediaminetetraacetic acid (EDTA), citric acid, cysteine, thioglycerol, thioglycolic acid, thiosorbitol, butylated Hydroxanisole, butylated hydroxyltoluene, and propyl gallate. Additionally, amino acids that are readily oxidized can be used as antioxidants, such as cysteine, methionine and tryptophan.

Examples of amino acids that may be included in the particle formulation include arginine, methionine, glycine, histidine, alanine, leucine, glutamic acid, iso-leucine, L-threonine, 2-phenylamine, valine, norvaline, proline, phenylalanine, tryptophan, serine, asparagine, cysteine, tyrosine, lysine and norleucine. Suitable amino acids include those that are readily oxidized, such as cysteine, methionine and tryptophan.

Examples of buffers that may be included in the particle formulation include, but are not limited to, citrate, histidine, succinate, phosphate, maleate, tris, acetate, carbohydrate, and gly-gly. Suitable buffers include citrate, histidine, succinate and tris.

Examples of inorganic compounds that may be included in the particle formulation include, but are not limited to, NaCl, Na2SO4, NaHCO3, KCl, KH2PO4, CaCl2 and MgCl2.

The particle formulation may also include other stabilizers/excipients such as surfactants and salts. Examples of surfactants include, but are not limited to, polysorbate 20, polysorbate 80, PLURONIC® (BASF Corporation, Mount Olive, NJ), F68, and sodium dodecyl sulfate (SDS). Examples of salts include, but are not limited to, sodium chloride, calcium chloride and magnesium chloride.

The particles are typically sized so that they can be delivered via an implantable osmotic delivery device. The uniform shape and size of the particles typically allows to provide a sustained and uniform rate of release from such delivery devices; However, particle formulations with non-normal particle size distribution profiles may also be used. For example, in a typical implantable osmotic delivery device having a delivery orifice, the particle size is less than about 30% of the diameter of the delivery orifice, more preferably less than about 20%, and even more preferably less than about 10%. In embodiments of the particle formulation for use with an osmotic delivery system, the delivery orifice diameter of the implant is about 0.5 mm, and the particle size can be, for example, less than about 150 microns to about 50 microns. In embodiments of the particle formulation for use with an osmotic delivery system, the delivery orifice diameter of the implant is about 0.1 mm, and the particle size can be, for example, less than about 30 microns to about 10 microns. In one embodiment, the orifice is about 0.25 mm (250 microns) and the particle size is from about 2 microns to about 5 microns.

Those skilled in the art will recognize that particle populations are subject to the principle of particle size distribution. A widely used, art-recognized method of describing particle size distribution includes, for example, an average diameter and a D value, such as a D50 value, to represent the average diameter of a range of particle sizes for a given sample. Commonly used.

The particles of the particle formulation are from about 2 microns to about 150 microns in diameter, e.g., less than 150 microns in diameter, less than 100 microns in diameter, less than 50 microns in diameter, less than 30 microns in diameter, less than 10 microns in diameter, less than 10 microns in diameter. It is less than 5 microns and about 2 microns in diameter. Preferably, the particles are from about 2 microns to about 50 microns in diameter.

The particles of the particle formulation comprising the isolated amylin analog polypeptide have an average diameter of from about 0.3 microns to about 150 microns. The particles of the particle formulation comprising the isolated amylin analog polypeptide have an average diameter of from about 2 microns to about 150 microns, e.g., less than 150 microns, less than 100 microns in average diameter, less than 50 microns in average diameter, average diameter It has an average diameter of less than 30 microns, an average diameter of less than 10 microns, an average diameter of less than 5 microns, and an average diameter of about 2 microns. In some embodiments, the particles have an average diameter of from about 0.3 microns to about 50 microns, such as from about 2 microns to about 50 microns. In some embodiments, the particles have an average diameter of from 0.3 microns to 50 microns, such as from about 2 microns to about 50 microns, wherein each particle is less than about 50 microns in diameter.

Typically, the particles of the particle formulation, when incorporated into a suspension vehicle, do not settle at delivery temperature in less than about 3 months, preferably in less than about 6 months, and more preferably in less than about 12 months. More preferably, it does not settle in less than about 24 months at the delivery temperature, and most preferably it does not settle in less than about 36 months at the delivery temperature. Suspension vehicles typically have a viscosity of from about 5,000 to about 30,000 poise, preferably from about 8,000 to about 25,000 poise, more preferably from about 10,000 to about 20,000 poise. In one embodiment, the suspension vehicle has a viscosity of about 15,000 poise, plus or minus about 3,000 poise. Generally speaking, smaller particles tend to have a slower sedimentation rate in viscous suspension vehicles than larger particles. Accordingly, micron- to nano-sized particles are typically preferred. In viscous suspension formulations, about 2 microns to about 7 microns particles of the present invention will not settle for at least 20 years at room temperature based on simulation modeling studies. In an embodiment of the particle formulation of the present invention, for use in an implantable osmotic delivery device, a particle size of less than about 50 microns, more preferably less than about 10 microns, more preferably in the range of about 2 microns to about 7 microns. include

In summary, the disclosed polypeptides, or pharmaceutically acceptable salts thereof, are formulated as a dry powder of solid state particles, which preserves the maximum chemical and biological stability of the drug. The particles provide long-term storage stability at high temperatures, thus making it possible to deliver stable and biologically effective drugs to a subject for extended periods of time. The particles are suspended in a suspension vehicle for administration to a patient.

Particle Suspension in Vehicle

In one aspect, the suspension vehicle provides a stable environment in which the drug particle formulation is dispersed. The drug particle formulation is chemically and physically stable (as described above) in a suspension vehicle. Suspension vehicles typically include one or more polymers and one or more solvents that form a solution of sufficient viscosity to uniformly suspend the drug-containing particles. Suspension vehicles may include additional ingredients including, but not limited to, surfactants, antioxidants, and/or other compounds soluble in the vehicle.

The viscosity of the suspension vehicle is typically sufficient to prevent settling of the drug particle formulation during storage and for use in delivery methods, eg, implantable, osmotic delivery devices. Suspension vehicles are biodegradable in that the suspension vehicle disintegrates or degrades over time in response to the biological environment, however, the drug particles dissolve in the biological environment and the active pharmaceutical ingredient (ie, drug) in the particles is absorbed.

In an embodiment, the suspension vehicle is a "single-phase" suspension vehicle that is a solid, semi-solid or liquid homogeneous system that is physically and chemically uniform throughout.

The solvent in which the polymer is dissolved can affect the characteristics of the suspension formulation, such as the behavior of the drug particle formulation during storage. The solvent may be selected in combination with the polymer such that the resulting suspension vehicle exhibits phase separation upon contact with an aqueous environment. In some embodiments of the invention, the solvent may be selected in combination with the polymer such that the resulting suspension vehicle exhibits phase separation upon contact with an aqueous environment having approximately less than 10% water.

The solvent may be an acceptable solvent that is immiscible with water. The solvent may also be selected such that the polymer is soluble in the solvent at high concentrations, such as greater than about 30% polymer concentration. Examples of solvents useful in the practice of the present invention are lauryl alcohol, benzyl benzoate, benzyl alcohol, lauryl lactate, decanol (also called decyl alcohol), ethyl hexyl lactate, and long chain (C8 to C24) aliphatic alcohols. , esters or mixtures thereof. The solvent used in the suspension vehicle may be "dry" in that it has a low water content. Preferred solvents for use in the formulation of suspension vehicles include lauryl lactate, lauryl alcohol, benzyl benzoate, and mixtures thereof.

Examples of polymers for formulation of suspension vehicles of the present invention include polyesters (e.g., polylactic acid and polylactic polyglycolic acid), pyrrolidone (e.g., polyvinylpi having a molecular weight ranging from approximately 2,000 to approximately 1,000,000). rolidone), esters or ethers of unsaturated alcohols (eg vinyl acetate), polyoxyethylenepolyoxypropylene block copolymers, or mixtures thereof. Polyvinylpyrrolidone can be characterized by its K-value (eg, K-17), which is the viscosity index. In one embodiment, the polymer is polyvinylpyrrolidone having a molecular weight of 2,000 to 1,000,000. In a preferred embodiment, the polymer is polyvinylpyrrolidone K-17 (typically with an approximate average molecular weight in the range of 7,900 to 10,800). The polymers used in the suspension vehicle may comprise one or more different polymers, or may comprise different grades of a single polymer. The polymer used in the suspension vehicle may also be dry or may have a low moisture content.

Generally speaking, suspension vehicles for use in the present invention may differ in composition based on the desired performance characteristics. In one embodiment, the suspension vehicle may comprise from about 40% to about 80% by weight of the polymer(s) and from about 20% to about 60% by weight of the solvent(s). A preferred embodiment of a suspension vehicle comprises a vehicle formed of polymer(s) and solvent(s) combined in the following ratios: from about 25% solvent to about 75% polymer; from about 50 weight percent solvent to about 50 weight percent polymer; from about 75 wt % solvent to about 25 wt % polymer. Thus, in some embodiments, the suspension vehicle may comprise selected ingredients, and in other embodiments consist essentially of selected ingredients.

Suspension vehicles may exhibit Newtonian behavior. Suspension vehicles are typically formulated to provide a viscosity that maintains a uniform dispersion of the particle formulation for a predetermined period of time. This facilitates the preparation of suspension formulations tailored to provide controlled delivery of the drug contained in the drug particle formulation. The viscosity of the suspension vehicle may vary depending on the intended use, the size and type of particle formulation, and the loading of the particle formulation in the suspension vehicle. The viscosity of the suspension vehicle can be varied by changing the type or relative amount of solvent or polymer used.

The suspension vehicle may have a viscosity ranging from about 100 poise to about 1,000,000 poise, preferably from about 1,000 poise to about 100,000 poise. In a preferred embodiment, the suspension vehicle typically has a viscosity of from about 5,000 to about 30,000 poise at 33° C., preferably from about 8,000 to about 25,000 poise, more preferably from about 10,000 to about 20,000 poise. In one embodiment, the suspension vehicle has a viscosity of about 15,000 poise at 33°C, plus or minus about 3,000 poise. Viscosity can be measured at 33° C. using a parallel plate rheometer at a shear rate of 10-4/sec.

Suspension vehicles may exhibit phase separation when contacted with an aqueous environment; However, typically suspension vehicles exhibit substantially no phase separation as a function of temperature. For example, at temperatures ranging from approximately 0° C. to approximately 70° C. and upon cycling temperature, such as from 4° C. to 37° C. to 4° C., the suspension vehicle typically exhibits no phase separation.

Suspension vehicles can be prepared by combining the polymer and solvent under dry conditions, such as in a dry box. The polymer and solvent can be combined at an elevated temperature, such as from about 40° C. to about 70° C., allowing it to liquefy and form a single phase. The ingredients may be compounded under vacuum to remove air bubbles generated from the dry ingredients. The ingredients may be combined using a conventional mixer set at a speed of approximately 40 rpm, such as a double helix blade or similar mixer. However, higher speeds may be used to mix the ingredients. Once a liquid solution of the ingredients is achieved, the suspension vehicle can be cooled to room temperature. Differential scanning calorimetry (DSC) can be used to confirm that the suspension vehicle is single phase. Additionally, the components of the vehicle (eg, solvents and/or polymers) can be substantially free of peroxide (eg, by treatment with methionine; see, eg, US Patent Publication No. 2007-0027105) peroxide. can be treated to reduce or substantially eliminate.

The drug particle formulation is added to a suspension vehicle to form a suspension formulation. In some embodiments, the suspension formulation may comprise a drug particle formulation and a suspension vehicle, and in other embodiments, it consists essentially of the drug particle formulation and the suspension vehicle.

Suspension formulations can be prepared by dispersing the steric formulation in a suspension vehicle. The suspension vehicle is heated and the particle formulation can be added to the suspension vehicle under dry conditions. The ingredients may be mixed under vacuum at an elevated temperature, such as from about 40°C to about 70°C. The ingredients may be mixed at a sufficient speed, such as from about 40 rpm to about 120 rpm, for a sufficient amount of time, such as about 15 minutes, to achieve a uniform dispersion of the three-dimensional formulation in a suspension vehicle. The mixer may be a double helix blade or other suitable mixer. The resulting mixture is removed from the mixer, sealed in a dry container, to prevent water from contaminating the suspension formulation, and at room temperature prior to further use, e.g., loading into an implantable, drug delivery device, unit dose container or multi-dose container. is cooled with

Suspension formulations typically have a total moisture content of less than about 10% by weight, preferably less than about 5% by weight, and more preferably less than about 4% by weight.

In a preferred embodiment, the suspension formulation of the present invention is substantially homogeneous and flowable to provide for delivery of the drug particle formulation from the osmotic delivery device to the subject.

In summary, the components of the suspension vehicle provide biocompatibility. The components of the suspension vehicle provide suitable chemical-physical properties to form a stable suspension of the drug particle formulation. These properties include, but are not limited to: the viscosity of the suspension; purity of vehicle; residual moisture in the vehicle; the density of the vehicle; compatibility with dry powder; compatibility with implantable devices; molecular weight of the polymer; stability of the vehicle; and the hydrophobicity and hydrophilicity of the vehicle. These properties can be manipulated and controlled, for example, by modification of the vehicle composition and manipulation of the component ratios used in the suspension vehicle.

The suspension formulations described herein can be used in implantable, osmotic delivery devices that provide zero-order, continuous, controlled and sustained delivery of a compound over an extended period of time, such as weeks, months, or up to about a year or more. there is. Such implantable osmotic delivery devices are typically capable of delivering a suspension formulation comprising a drug at a desired flow rate over a desired period of time. The suspension formulation can be loaded into an implantable, osmotic delivery device by conventional techniques.

implantable delivery

The dose and rate of delivery can be selected to achieve a desired blood concentration of the drug, which is generally within a half-life of less than about 6, in a subject after device implantation. The blood concentration of the drug provides the optimal therapeutic effect of the drug, while avoiding the unwanted side effects that can be induced by excessive drug concentration, while at the same time peak and trough that can induce side effects related to the peak or trough plasma concentration of the drug. is chosen to avoid

Implantable, osmotic delivery devices typically include a reservoir having at least one orifice through which the suspension formulation is delivered. The suspension formulation may be stored in a reservoir. In a preferred embodiment, the implantable drug delivery device is an osmotic delivery device, wherein delivery of the drug is osmotically directed. Some osmotic delivery devices and some of their components, such as DUROS® delivery devices or similar devices, have been described (see, eg, US Pat. Nos. 5,609,885; 5,728,396; 5,985,305; 5,997,527; 6,113,938). 6,132,420; 6,156,331; 6,217,906; 6,261,584; 6,270,787; 6,287,295; 6,375,978; 6,395,292; 6,508,808; 6,544,252; 6,6,68,268; 6,923,800; 6,939,556; 6,976,981; 6,997,922; 7,014,636; 7,207,982; and 7,112,335; 7,163,688; US Patent Publication Nos. 2005/0175701, 2007/0281024, 2008/ 0091176 and 2009/0202608).

Osmotic delivery devices typically consist of an osmotic engine, a piston, and a cylindrical reservoir containing the drug formulation. The reservoir is capped at one end by a controlled rate, semipermeable membrane, and at the other end by a diffusion regulator through which the drug-containing suspension formulation is released from the drug reservoir. The piston separates the drug formulation from the osmotic engine and utilizes a seal to prevent water in the osmotic engine compartment from entering the drug reservoir. The diffusion regulator is designed with the drug formulation to prevent body fluids from entering the drug reservoir through the orifice.

The osmotic device releases the drug at a predetermined rate based on osmotic principles. The extracellular fluid enters the salt engine directly through the semipermeable membrane and the osmotic delivery device expands to drive the piston at a slow, uniform delivery rate. Movement of the piston exerts a force such that the drug formulation is released through the orifice or exits the port at a predetermined delivery rate. In one embodiment of the invention, the reservoir of the osmotic device is loaded with a suspension formulation, wherein the device is delivered at a predetermined, therapeutically effective delivery rate for an extended period of time (e.g., about 1, about 3, about 6, about 9, about 10, to about 12 months).

The rate of drug release from the osmotic delivery device typically provides the subject with a predetermined target dose of the drug, eg, a therapeutically effective daily dose delivered over the course of a day; That is, the rate of release of the drug from the device provides substantially steady state delivery of the drug at a therapeutic concentration to the subject.

Typically, for an osmotic delivery device, the volume of the advantageous formulation chamber containing the advantageous agent formulation is from about 100 μl to about 1000 μl, more preferably from about 120 μl to about 500 μl, more preferably from about 150 μl to about 200 μl.

Typically, osmotic delivery devices are implanted in a subject, eg, subdermally or subcutaneously, to provide subcutaneous drug delivery. The device(s) may be implanted subdermally or subcutaneously in one or both arms (eg, inside, outside, or behind the upper arm) or in the abdomen. A preferred location in the abdominal region is under the abdominal skin in the region extending below the ribs and above the belt line. To provide multiple locations for implantation of one or more osmotic delivery devices in the abdomen, the abdominal wall may be divided into quarters as follows: an upper right quadrant extending at least 2-3 centimeters below the right rib, for example , at least about 5-8 centimeters below the right rib, and at least 2-3 centimeters to the right of the midline, eg, at least about 5-8 centimeters to the right of the midline; the lower right quadrant extending at least 2-3 centimeters above the belt line, for example at least about 5 to 8 centimeters above the belt line, and at least 2-3 centimeters to the right of the center line, for example at least about 5 to 8 centimeters to the right of the center line centimeter; upper left extending at least 2-3 centimeters below the left rib, for example at least about 5-8 centimeters below the left rib, and at least 2-3 centimeters to the left of the midline, for example at least about 5-8 centimeters to the left of the midline centimeter; and a lower left quadrant extending at least 2-3 centimeters above the belt line, for example at least about 5 to 8 centimeters above the belt line, and at least 2-3 centimeters to the left of the center line, for example at least about 5 to left of the center line 8 centimeters. This provides a number of available devices for implantation of one or more devices in more than one instance. Implantation and removal of the osmotic delivery device is typically performed by a healthcare professional using local anesthesia (eg, lidocaine).

Termination of treatment by removal of the osmotic delivery device from the subject is direct and provides the subject with an important advantage of interim interruption of drug delivery.

Preferably, the osmotic delivery device is a safety guarantee to prevent inappropriate overdose or bolus delivery of the drug in theoretical situations such as plugging or clogging of the outlet (diffusion regulator) to which the drug formulation is delivered. have a device In order to prevent inappropriate overdose or bolus delivery of drug, the pressure required to partially or fully remove or drain the diffusion regulator from the reservoir is necessary to partially or fully remove or evacuate the semipermeable membrane to such an extent that it is necessary to remove the reservoir pressure. The osmotic delivery device is designed and constructed to exceed the pressure. In this scenario, pressure is built in the device until it pushes the semipermeable membrane outward at the other end, thereby releasing osmotic pressure. The osmotic delivery device then becomes static and no longer delivers the drug formulation under the condition that the piston is in sealing relationship with the reservoir.

Suspension formulations can also be used in infusion pumps, such as ALZET® (DURECT Corporation, Cupertino, CA) osmotic pumps, which are miniature, infusion pumps for continuous dosing of laboratory animals (eg, mice and rats). there is.

Example

The following examples, together with a complete disclosure and description of how to practice the invention, are presented to provide those skilled in the art, and are not intended to limit the scope of what the inventors regard as the invention. Efforts have been made to ensure accuracy with respect to numbers used (eg, amounts, concentrations, and percentage changes), but some experimental errors and deviations should be accounted for. Unless otherwise indicated, temperature is in degrees Celsius and pressure is at or near atmospheric.

Example 1 Generation of Amylin Analog Polypeptides

Amylin analog polypeptides of the invention as provided in Table 3 are N-[(dimethylamino)-1H-1,2,3-triazolo-[ in N,N-dimethylformamide (DMF)) 4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (HATU) or 2-(6-chloro-1-H-benzotriazol-1-yl) Prelude peptide synthesizer (Protein Technologies Inc.) by solid-phase method using Fmoc power with -1,1,3,3-tetramethylaluminum hexafluorophosphate (HCTU) activation (5-fold molar excess relative to amino acid). , Tucson, Arizona)), and N'N-diisopropylethylamine (DIEA) was used as the base. A 20% piperidine/DMF solution was used for Fmoc deprotection. The resin used was Rink Amide MBHA LL (Novabiochem) using a load of (0.30 to 0.40) mmol/g on a (20 to 400) μmol scale.

Final deprotection and cleavage of the peptide from the solid support was performed by treatment of the resin with (92.5% TFA, 2.5% phenol, 2.5% water and 2.5% triisopropylsilane) for 2-3 hours. The cleaved peptide was precipitated using cold diethyl ether. The diethyl ether was decanted and the solid was triturated again with diethyl ether and pelleted by centrifugation. The crude solid was then dissolved in a 1:1 solution of ACN/water, 0.01% TFA. Disulfide bridge formation was obtained through the addition of a solution of iodine/acetic acid (35 mg/ml) to the solution of the crude product until the color of the solution continued to change to amber. The reaction solution was stirred until analysis via LC/MS indicated completion of the reaction. A 2% solution of ascorbic acid in _{H 2} O was added until the solution turned clear. The final crude product solution was lyophilized for final purification.

The lyophilized solid was redissolved in a 1:1 solution of acetonitrile/water with 0.1% TFA (10-15 mL), 0.1% over 30-45 minutes at a flow rate of 30 mL/min, λ - 215 nm. Purification via reverse phase on a Waters XBridge™ BEH 130, CIS, 10pm, 130 Å, 30×250 mm ID column using a 30 gradient within the range of 5 to 75% acetonitrile/water with TFA.

Example 2: Purification and characterization of amylin analog polypeptides, i.e., linear polypeptides, without any lipophilic substituents and optional spacers

The purified product was lyophilized and analyzed by ESI-LC/MS and analytical HPLC and proved to be pure (>98%). All mass results were in agreement with the calculated values.

UV provided by C18 HPLC and LC/MS analysis (Acquity SQD Waters Corp, Milford, MA) and dual absorbance signals at 215 nm and 280 nm using either Method A, Method B, Method C or Method D Characterization of the peptide analogs was performed by detection.

Method A, LC/MS conditions: Phenomenex UPLC Aeris™ Peptide XB Cl8 35 column, 1.7 pm, 2.1×100 mm or ACQUiTY BEH300 or BEH130 CT8 column, 1.77 pm. 2.1×100 mm using 5 to 65% acetonitrile/water with 0.05% TFA over 30 minutes at a flow rate of 0.5 ml/min, λ - 215 nm, 280 nm.

Method B, C18 HPLC conditions: Acquity BEH130, C18 column, 1.7 μm, 0.05% TFA over 30 min, flow rate 0.5 ml/min, l 215 nm, 25° C. at l 280 nm, 5-65% acetonitrile/water, 1.7 μm, UPLC analysis was performed on a 100×2.10 mm column.

Method C, UPLC conditions: Acquity BEH130, C18 column, 1.7 μm, 100 with 0.05% TFA over 20 min, flow rate 0.5 ml/min, l 215 nm, l 280 nm at 25° C., 5-65% acetonitrile/water UPLC analysis was performed on a x 2.10 mm column.

Method D, UPLC conditions: Acquity BEH130, C18 column, 1.7 μm, 100 with 0.05% TFA over 10 min, flow rate 0.5 mL/min, l 215 nm, l 280 nm at 25° C., 5-65% acetonitrile/water UPLC analysis was performed on a x 2.10 mm column. 5.0 μl of the sample was injected using the PLNO (partial loop with needle over-fill) injection method.

Table 3 provides exemplary amylin analog polypeptides of the present disclosure.

Polypeptide analogs lacking lipophilic substituents and optional spacers are sometimes referred to herein as “linear polypeptides”. Polypeptide analogs having at least one covalently linked lipophilic substituent and an optional spacer are sometimes referred to herein as "conjugated polypeptides".

Example 3: Synthesis of Amylin Analog Polypeptide Intermediate

Synthesis of polypeptides with modifications at the D- Lys16 or L-Lys16 positions, for example (Compounds B2, A129, B4 and B8)

Upon completion of the linear polypeptide synthesis as described in Example 1, the resin was washed with dichloromethane (DCM) and dried under vacuum for 30 minutes. For analogs containing an Alloc-protecting group, their removal was facilitated via a solution _{of Pd(PPh 3} ) _{3 in} (chloroform/acetic acid/n-methyl-morpholine, 37:2:1). The resulting deprotection resin was washed with 2% diethyldithiodicarbamate sodium trihydrate/DMF (6×30 sec), 2% DIEA/DMF (6×30 sec), and finally DMF (6×30 sec). did. Extension of the spacer region was performed in a stepwise manner by manual addition of each building block under pre-activation conditions. To 1 ml of a 200 mmol solution of Fmoc-γGlu-(OH)-OtBu in DMF, 0.5 ml of DIEA (800 mmol), followed by 0.5 ml of HCTU (400 mmol) was added, and the resulting reaction solution was stirred for 5 minutes. However, it was added to the deprotected residues on the linear sequence. The reaction mixture was stirred under nitrogen for 30 minutes. The resin was decanted and washed with DMF (6x30 sec). Subsequent removal of the Fmoc protecting group was facilitated with 20% piperidine/DMF followed by a final dilution with DMF (6×30 sec). Final deprotection and cleavage of the peptide from the solid support was performed by treatment of the resin with (95% TFA, 2% water, 2% thioanisole and 1% triisopropylsilane) for 2-3 hours. Each intermediate was isolated by incorporation of each building block using the methods described above and characterized by HPLC/MS.

Synthesis of polypeptides with modifications at the N-terminus, for example (A130, B22 and B23)

The synthesis of the linear sequence was performed as described in Example 1. Addition of the albumin-binding moiety was facilitated by removal of the Fmoc-protecting group at the N-terminus via a 20% solution of piperidine/DMF. The resin was washed with DMF and the incorporation of side chain building blocks was performed in a stepwise manner under pre-activation conditions. To 1 ml of a 200 mmol solution of Fmoc-γGlu-(OH)-OtBu in DMF was added 0.5 ml of DIEA (800 mmol) followed by 0.5 ml of HCTU (400 mmol). The resulting reaction solution was stirred for 5 minutes, which was then added to the deprotected linear sequence. The reaction mixture was stirred under nitrogen for 30 minutes. The resin was decanted and washed with DMF (6x30 sec). Final deprotection and cleavage of the peptide from the solid support was performed by treatment of the resin with (95% TFA, 2% water, 2% thioanisole and 1% triisopropylsilane) for 2-3 hours. Each intermediate was isolated and characterized via HPLC/MS. Chemical data for both internally modified polypeptides and N-terminally modified polypeptide intermediates are reported in Table 5 below.

Example 4: Covalent attachment of lipophilic substituents and optional spacers to an amylin analog polypeptide, i.e., conversion of a linear polypeptide to a conjugated polypeptide

Synthesis of an amylin analog polypeptide conjugated with one or more albumin-binding lipophilic substituents and optional spacers was performed with a modification of the synthetic method described in Example 1.

Upon completion of the linear polypeptide synthesis as described in Example 1, the resin was washed with dichloromethane (DCM) and dried under vacuum for 30 minutes. For analogs containing alloc-protecting groups, their removal was obtained via a solution _{of Pd(PPh 3} ) _{3 in} (chloroform/acetic acid/n-methyl-morpholine, 37:2:1). For analogs containing BOC-Lys(Fmoc)-OH, the Fmoc protecting group was removed using 20% piperidine/DMF. The resulting deprotected resin was washed with DMF (6×30 sec). Next, extension of the spacer region was performed in a stepwise manner by manual addition of each building block under pre-activation conditions. Addition of lipophilic substituents (also referred to as “acyl chains”) was performed under normal SPPS conditions without a prior activation step. Final deprotection and cleavage of the peptide from the solid support was performed by treatment of the resin with (95% TFA, 2% water, 2% thioanisole and 1% triisopropylsilane) for 2-3 hours. The cleaved peptide was precipitated using cold diethyl ether. The diethyl ether was decanted and the solid was triturated again with diethyl ether and pelleted by centrifugation.

The crude product was then dissolved in a solution of ACN/H2O, 0.1% TFA. A solution of iodine/acetic acid (35 mg/ml) was added to each solution of the crude peptide product until the color of the solution continued to change to amber. The reaction solution was stirred until analysis via LC/MS indicated that the desired disulfide bridge was formed. A 2% solution of ascorbic acid in _{H 2} O was added to the reaction solution until the solution turned clear. The solution was frozen and lyophilized. A tablet was obtained through the method described in Example 1.

A typical synthesis of conjugated peptides is described. Synthesis of A109 : The synthesis of the linear sequence was performed as described in Example 1. Addition of the albumin-binding moiety was facilitated by removal of the Fmoc-protecting group at the N-terminus via a 20% solution of piperidine/DMF. The resin was then washed with DMF, and the incorporation of side chain building blocks was performed in a stepwise manner under pre-activation conditions. To 1 ml of a 200 mmol solution of Fmoc-γGlu-(OH)-OtBu in DMF was added 0.5 ml of DIEA (800 mmol) followed by 0.5 ml of HCTU (400 mmol). The resulting reaction solution was stirred for 5 minutes, which was then added to the deprotected linear sequence. The reaction mixture was stirred under nitrogen for 30 minutes. The resin was then decanted and washed with DMF (6x30 sec). The Fmoc-protecting group of γGlu was removed with 20% piperidine/DMF. It was then combined with octadecanedione (C18) (200 mmol) in DMF using HATU (400 mmol) and DIEA (800 mmol) under normal solid phase conditions.

Cleavage was obtained using a 95% TFA/2% water/2% thioanisole/1% TIPS solution. The crude product was then _{dissolved in a solution of ACN/H 2} O, 0.1% TFA. A solution of iodine/acetic acid (35 mg/ml) was added to the solution of the crude peptide product until the color of the solution changed continuously to amber. The reaction solution was stirred until analysis via LC/MS indicated that the desired disulfide bridge was formed. A 2% solution of ascorbic acid in _{H 2} O was added to the reaction solution until the solution turned clear.

Lyophilization of the crude product gave an off-white solid, which was purified by the method described in Example 2.

Example 5: Stability of Amylin Analog Polypeptides

Several amylin analog polypeptides described herein were tested for stability as trifluoro acetate salts in 1 mg/ml solution in DMSO (ie, organosulfur solvent) or in aqueous (ie, deionized water). These analog polypeptides were incubated at 37° C. and samples were withdrawn at various time intervals and analyzed by LC/MS and HPLC for determination of the purity and mass of the parent peptide and the extent of any degradation products. The purity results of these assays are shown in Tables 6A and 6B and are considered indicative of stability.

Example 6: Solubility of Amylin Analog Polypeptides

The analog polypeptides described herein were tested for solubility in saline 20% DMSO in water (ie, bioassay buffer) or in aqueous (deionized water) at room temperature. The samples were visually examined for clarity, any appearance of turbidity or haze. Table 7 shows the results of this analysis.

Example 7: Functional Assay: Human Calcitonin Receptor and Amylin 3 Receptor

Activation of the human calcitonin receptor (hCTR), or human amylin 3 receptor (hAMY3R), results in an increase in cyclic adenosine modophosphate (cAMP) in cells. In the presence of the non-specific cAMP/cGMP phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX), cAMP accumulating can be measured in vitro using conventional detection methods. Thus, the fitted dose-response curves for cAMP accumulation can be used to estimate the in vitro potency (pEC50) for peptides activating each of these receptors.

Cell, culture and cAMP assays

HEK293-CNG stably expressing human calcitonin receptor (hCTR) or co-expressing human calcitonin receptor and human receptor activity modifying protein 3 (hAMY3R) (Codex Biosolutions #CB-80200-258 and #CB-80-200-271, respectively) Cells were treated with 90% DMEM, 10% FBS, 250 μg/ml G418 and 1 μg/ml puromycin (hCTR cells), or 90% DMEM, 10% FBS, 250 μg/ml G418, 1 μg/ml puromycin, 150 Grown in μg/ml hygromycin B (hAMY3R cells). Cells were transferred in growth medium for up to 10 passages prior to testing.

On the day of analysis, cells expressing hCTR or hAMY3R are counted and 1× HBSS, 5 mM HEPES, 0.5 mM IBMX, and 0.1% bovine serum albumin (BSA) with 0%, 0.1% or 4% human serum albumin (HSA) ) or in white 384-well OptiPlates (PerkinElmer #6007299) in 5 mcL stimulation buffer consisting of 1× HBSS, 5 mM HEPES, 0.5 mM IBMX, and 0.1% casein at 500 cells per well.

The peptides were serially diluted in the same buffer as above for each given assay condition. Two assay control solutions consisting of either 50 mcM forskolin (cAMP system maximum) or assay buffer alone (cAMP system minimum) were also prepared in appropriate stimulation buffer. Five milliliters of each peptide concentration or assay control was added to the wells in triplicate and incubated for 30 minutes at room temperature. During this incubation step, 4x Europium labeled cAMP tracer solution and 4x Ulight® anti-cAMP solution (consisting of anti-cAMP monoclonal antibody labeled with Ulight™ dye) were added to the manufacturer's protocol (PerkinElmer LANCE Ultra cAMP kit). prepared according to After this incubation, 5mcL Europium labeled cAMP and 5mcL Ulight anti-cAMP antibody were added to the wells. The plate was covered with an adhesive cover to prevent evaporation and incubated in the dark for 60 min at room temperature. Plates were read on an Envision fluorescence plate reader (PerkinElmer).

data analysis

The test values in Excel were initially normalized to the mean value of the forskolin-induced cAMP system maximum and system minimum using the formula: (test value - mean system minimum)/(mean system maximum - mean system minimum) * 100. Normalized test values represent the reference corrected percentage of the system maximal cAMP response induced by forskolin. Normalized data were analyzed from triplicate trials and used to estimate EC50 for each peptide on each receptor. Data were fitted in GraphPad Prism software (v7.04) using a 4-parameter logistic curve model: Y=bottom ∀+ (top-bottom)/(1+10^((LogEC50-X))). Hill slope was limited to 1.0. EC50 values were converted to pEC50 values using the formula: pEC50 = - Log (EC50).

Data Interpretation

For a given receptor, in vitro potency estimates (pEC50) in the absence of HSA are similar for all peptides, as they reflect albumin independent binding (free peptide) values (0% HSA, Table 8). However, in the presence of albumin (BSA or HSA), potency measurements across acylated peptides are not distinctly similar (Table 8). This is due to the variability of the albumin-acylpeptide binding efficiency which is collectively dependent on the amino acid sequence, the acyl binding motif, the site of attachment, and the length of the linker engineered into each peptide. However, for a given junction polypeptide, a decrease in potency in the presence of HSA (decreased pEC50 values) is indicative of an albumin-peptide interaction quantitatively (Table 8, FIGS. 2A and 2B ). In contrast, the linear polypeptides (pramlintide, hcalcitonin) were not affected by the presence of HSA (Table 8, Figure 2C and Figure 2D), reflecting their poor albumin binding efficiency.

Example 8: Intravenous Infusion of “Linear” (ie, Non-acylated) Amylin Analog Polypeptides: A Pharmacokinetic Study to Assess Clearance (CL) from the Kidney of Linear Amylin Analog Polypeptides

Peptides were dissolved in sterile saline and administered as a 3-hour intravenous infusion to non-fasting male Sprague-Dawley rats (n=3 per group) via femoral vein cannula at a final dose of 0.100 mg/kg. The formulation was administered at a rate of 1.67 ml/kg/h. At 0.25, 0.5, 1, 2, 3, 3.17, 3.33, 3.5, 4, 4.5, 5 and 6 hours after initiation of infusion (long method) or at 1, 1.5, 2, 2.5 and 3 hours after initiation of infusion (steady state) Screening Method) Blood samples (approximately 250 μl) were collected for pharmacokinetic analysis via jugular vein cannula. All samples were collected in microtainer tubes containing K2EDTA as anticoagulant and 25 μl of protease inhibitor cocktail. Plasma was prepared by centrifugation and stored at -80°C until analysis. The results of this analysis are shown in Table 9, some of which are illustrated in Figures 3A-3B.

Example 9: Conjugated (i.e., acylated) Amylin Analog Polypeptides to Assess Clearance (CL) from the Kidney of Conjugated Amylin Analog Polypeptides: Intravenous Injection of a Pharmacokinetic Study

Peptides were dissolved in sterile saline and administered as a 1-hour intravenous infusion to non-fasting male Sprague-Dawley rats (n=3 per group) via femoral vein cannula at a final dose of 0.033 mg/kg. The formulation was administered at a rate of 1.67 ml/kg/h. Jugular vein at 0.25, 0.5, 0.75, 1, 1.17, 1.33, 1.5, 2, 4, 6, 8, 24, 30 and 48 hours after infusion into microtainer tubes containing K2EDTA as anticoagulant and 25 μl of protease inhibitor cocktail Blood samples (approximately 250 μl) were collected via cannula for pharmacokinetic analysis. Plasma was prepared by centrifugation and stored at -80°C until analysis. The results of this analysis are shown in Table 9, some of which are illustrated in Figures 3A-3B.

Example 10: Subcutaneous Injection: Pharmacokinetic Study to Assess Clearance (CL) from the Kidney of Amylin Analog Polypeptides

Peptide was dissolved in sterile saline and as a 1-hour subcutaneous injection into non-fasting male Sprague-Dawley rats (n=3 per group) via a cannula placed into the subcutaneous space between the scapula at a final dose of 0.033 mg/kg. administered. The formulation was administered at a rate of 0.145 ml/h/kg. Pharmacokinetics via jugular cannula at 0.25, 0.5, 1, 1.5, 2, 4, 6, 8, 24, 30 and 48 hours after initiation of infusion into microtainer tubes containing K2EDTA as anticoagulant and 25 μl of protease inhibitor cocktail. A blood sample (approximately 250 μl) was collected for analysis. Plasma was prepared by centrifugation and stored at -80°C until analysis. Table 9 shows the results of this analysis.

Example 11: Subcutaneous Bolus Injection: A Pharmacokinetic Study to Assess Clearance (CL) from the Kidney of Amylin Analog Polypeptides

Peptides were dissolved in sterile saline and administered to non-fasting male Sprague-Dawley rats (n=3 per group) via a single bolus injection into the subcutaneous space between the scapula at a dose of 0.3 mg/kg. Pharmacokinetic analysis via jugular vein cannula at 0.083, 0.167, 0.25, 0.5, 1, 2, 4, 8, 24, 30 and 48 hours post-dose into microtainer tubes containing K2EDTA as anticoagulant and 25 μl of protease inhibitor cocktail. A blood sample (approximately 250 μl) was collected for Plasma was prepared by centrifugation and stored at -80°C until analysis. Table 9 shows the results of this analysis.

Example 12: Method of Preparing Plasma Samples for Pharmacokinetic Studies

protein precipitation

A 60 μl aliquot of each plasma sample was placed in a 96-well plate. 6 μl of 0.5% Tween-20 was added to each well. The plates were then stirred and mixed at 1200 rpm for 10 minutes before 180 μl of 0.1% TFA in 2:1 ethanol:acetonitrile containing the appropriate internal standard was added to each well. Plates were stirred and mixed at 1300 rpm for 5 minutes and centrifuged at 2844×g for 10 minutes. The supernatant (180 μl) was placed in a clean 96-well plate and evaporated at 45° C. under a stream of nitrogen. The residue was reconstituted in 80 μl of 20% acetonitrile (aq) containing 0.1% formic acid.

solid phase extraction

A 60 mL aliquot of each plasma sample was diluted with 180 mL of 10 mM ammonium acetate, pH 6.8, containing the appropriate internal standard, in Oasis WCX microElution plates (Waters Corporation) pre-conditioned with 200 mL of methanol and 200 mL of deionized water. , Milford, Massachusetts). Samples were washed with 200 mL of 5% ammonium hydroxide (aq) followed by 200 mL of 20% acetonitrile in water. The analyte was eluted with 200 mL of 75:25 acetonitrile:5% formic acid in water. The eluent was dried under a stream of nitrogen. The residue was reconstituted in 80 μl of 20% acetonitrile (aq) containing 0.1% formic acid.

Example 13: LC/MS Quantification of Amylin Analog Polypeptides in Plasma

All calibration standards were prepared in control rat plasma containing K2EDTA and a protease inhibitor cocktail.

CTC HTS PAL Autoinjector (Leap, Calborough, NC), Agilent Infinity 1290 System with Column Oven (Palo Alto, CA), Valco Switch Valve (Houston, TX), and AB Sciex API 5600 Samples and standards were analyzed by TurboIonSpray™ UPLC-MS/MS using a system consisting of a TripleTOF™ or Sciex API 4000QTrap mass spectrometer (Framingham, MA). Samples were injected into a 2.1 x 50 mm reversed-phase C18 analytical column, typically a Waters CORTECS UPLC C18+, 1.6 μm (Waters Corporation, Milford, MA) or the like. Chromatographic separation was achieved using a gradient method using water (A) containing 0.1% formic acid and acetonitrile (B) containing 0.1% formic acid as mobile phases. Initial conditions consisted of 90% A and 10% B. Depending on the peptide, the organic component was increased to 95% B over a period of 3 to 4 minutes. A typical flow rate was 600 μl/min. The column temperature was kept constant at 40 or 50°C. Peptides were quantified by monitoring one or more product ions generated from multiple charged parent ions.

Example 14: In Vivo Efficacy of Amylin Analog Polypeptides Using Food Intake Inhibition in Rats

Rapid food intake was measured continuously for 72 hours using a BioDAQ food monitoring system (Research Diets, New Brunswick, NJ) to determine the amount of food intake inhibition exhibited by these amylin analog polypeptides. Approximately 8 weeks old Long Evans rats were obtained. Rats were housed alone and acclimatized to a 45% high fat diet for at least 2 weeks prior to dosing. After 1 week of acclimatization, all rats were housed alone in BioDAQ cages (Research Diets, New Brunswick, NJ) and 12 h light/dark cycle at constant temperature (approximately 22° C.) and 30-70% relative humidity. (People from 7:00 am to 7:00 pm) were maintained. Rats were provided with ad libitum access to water and pellet chow (Research Diets D12451i, 45 kcal% fat, Research Diets, New Brunswick, NJ). All procedures were performed in accordance with the Animal Welfare Act, USDA regulations, and were approved by the Mispro Institutional Animal Care and Use Committee. Animals were randomized into treatment groups according to body weight (n=8 rats/group). Animals were dosed with specified concentrations of amylin analog polypeptide or vehicle control (saline) (SC bolus injection), dosed from 6:00 to 6:30 before light cycle by opening and closing the hopper, while animals were administered was administered. The hopper gate was opened and continuous data collection began immediately after dosing was completed. Data were analyzed using BioDAQ Viewer software (version 2.3.07) and filters were set if necessary to reduce noise in data associated with non-emotional behavior. All data are expressed as % inhibition from vehicle control and summarized as mean. Data were analyzed for statistical significance using Microsoft Excel (Raymond, Wash.) by two-sample t-test. A P-value < 0.05 was considered to indicate a significant difference between treatment groups. The % fast food intake from vehicle control results for amylin analog polypeptides are shown in Table 10.

Example 15: In vivo efficacy according to body weight change in LE DIO rats after 13 days

A chronic (13-day) in vivo dose-response efficacy study was performed in a rodent model for obesity (Long Evans (LE) diet-induced obesity (DIO) rats). Efficacy and persistence of amylin analog polypeptides on weight loss was studied. Male LE DIO rats were used (Envigo Laboratories, Inc., Indianapolis, IN), weaning was initiated and rats were fed a high fat diet (Teklad TD 95217, 40% kcal from fat, Harlan Laboratories, Wisconsin). based in Madison). Rats were 15-17 weeks old at the start of the study. Rats were housed 1 per cage, provided with a high fat diet (Harlan TD.95217, 4.3 kcal/g) and ad libitum access to water, from 5:00 am to 5:00 pm at 21° C. and 50% relative humidity. A 12-hour light/dark cycle was maintained until 0:00 and allowed to acclimatize for at least 10 days prior to surgery. All procedures were performed in accordance with the Animal Welfare Act, USDA regulations, and were approved by the Mispro Institutional Animal Care and Use Committee. Weight measurements were taken twice/week starting 3 days prior to surgery. Baseline fat mass and non-fat mass measurements were taken 3 days prior to the start of peptide injection using a QMR instrument (Echo Medical Systems, Houston, TX). Rats were randomized into various treatment groups (n=4 to 6 rats/group) according to their body weight fat mass percentage and/or body weight. Alzet mini-osmotic pumps (2 weeks; Model 2002, Durect Corporation, Cupertino, CA) were filled under sterile conditions with vehicle or peptide one day prior to surgery. On the day of surgery, the rats were anesthetized with isoflurane, and the dorsal skin surface was shaved and cleaned. Rats were injected SC with flunexin (2.5 mg/kg). A surgical incision of 1 to 2 cm was made between the scapula. Using blunt dissection, a 2 to 3 cm subcutaneous tunnel was created into which a sterile, filled, mini-osmotic pump was introduced. The skin opening was closed with a skin staple. Each rat was implanted with one or two osmotic pumps containing vehicle or peptide depending on their treatment group. Data were analyzed in Excel and/or Prism (GraphPad Software, Inc., La Jolla, CA) using one-way ANOVA to compare each group to the appropriate control group. A P-value < 0.05 was considered to indicate a significant difference between treatment groups. The mean weight loss (%) (ΔΔ) from baseline and vehicle controls from the 13 day study is shown in Table 9.

Example 16: Weight Loss Efficacy of Amylin Analog Polypeptides in Combination with Exenatide in LE DIO Rats

A long-term study was performed to determine the effectiveness and persistence of continuous administration of an amylin analog polypeptide in combination with exenatide (GLP-1 receptor agonist) on body weight after 27 days of treatment in LE DIO rats. Amylin analog polypeptide and/or exenatide (10 mcg/kg/d = ED50 for weight loss) or vehicle (20% DMSO in water) in 18-week-old male LE DIO rats (14 weeks on high fat diet) ( Two (2) Alzet osmotic mini-pumps containing one of the indicated doses (n=8 animals/treatment group) were implanted subcutaneously. Amylin analog polypeptides for which PK supports every other day dosing (eod) were administered by SC injection eod instead of mini pump administration. All other procedures were the same as described in the previous examples. Table 11 shows the mean weight loss (%) from baseline and vehicle control (ΔΔ) results from long-term combination studies with exenatide.

Example 17: Anti-diabetic Efficacy of Amylin Analog Polypeptides in Combination with Exenatide in ZDF Rats

To determine the antidiabetic effect of continuous administration of an amylin analog polypeptide in combination with exenatide on HbA1c (primary anti-diabetic parameter) after 27 days of treatment in Zucker Diabetic Fatty (ZDF) rats A long-term study was conducted to Male ZDF rats were obtained at 6 (6) weeks of age (Charles River, Raleigh, NC) and used in the study at 8 (8) weeks of age. When housed, rats are housed as one animal per cage with free access to Purina 5008 chow (Lab Diet, St. Louis, MO) and water, 5:00 am to pm at 21° C. and 50% relative humidity. A 12 h light/dark cycle was maintained until 5:00 h and allowed to acclimatize for nine (9) days prior to study initiation. Blood samples were taken as whole blood (-Day 3) via tail vein to measure glucose levels and HbA1c. ZDF rats were randomized into treatment groups (n=10/group) according to similar mean HbA1c and glucose. Alzet osmotic mini-pumps containing specified doses of amylin analog polypeptide and/or exenatide (10 mcg/kg/day) or vehicle (20% DMSO in water) (n=10 animals/treatment group) (two (2) pumps/animal) were implanted subcutaneously (SC). Amylin analog polypeptides for which PK supports every other day dosing (eod) were administered by SC injection eod instead of mini pump administration. All other procedures were the same as described in the previous examples. Blood samples were again taken on days 14 and 27 (end of study) to measure glucose levels and HbA1c. Final whole blood samples were collected by cardiac puncture under isoflurane anesthesia (day 27). HbA1c analysis was performed using a Carolina Chemistries CLC720i Clinical Chemistry Analyzer (Mindray Inc., Miwa, NY) with protocol and method parameters as described by the manufacturer. HbA1c results expressed as mean % change from baseline and vehicle control (ΔΔ) from long-term combination studies with exenatide are presented in Table 11.

While the invention has been described in conjunction with the detailed description thereof, the foregoing description is for the purpose of illustration and is not intended to limit the scope of the invention, which is defined solely by the scope of the appended claims. Other aspects, advantages and modifications are within the scope of the following claims.

Other embodiments

While the invention has been described in conjunction with the detailed description thereof, the foregoing description is for the purpose of illustration and is not intended to limit the scope of the invention, which is defined solely by the scope of the appended claims. Other aspects, advantages and modifications are within the scope of the following claims.

SEQUENCE LISTING <110> INTARCIA THERAPEUTICS, INC. <120> HUMAN AMYLIN ANALOG POLYPEPTIDES AND METHODS OF USE <130> IPA210506-US <140> PCT/US2019/055696 <141> 2019-10-10 <150> US 62/744,236 <151> 2018-10-11 <160> 318 <170> PatentIn version 3.5 <210> 1 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 1 His Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 2 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 2 Ile Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 3 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys(CO(CH2)4CO2H) <400> 3 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 4 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 4 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ser Asn Phe Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 5 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(gammaGlu-dpeg-dpeg-(gammaGlu)4-CO(CH2)14CH3) <400> 5 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 6 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Leu <400> 6 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 7 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 7 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Lys Thr Lys Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 8 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 8 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Lys Thr Tyr 35 <210> 9 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 9 Ser Cys Asn Thr Ala Ser Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 10 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (17)..(17) <223> Lys(gammaGlu-CO(CH2)14CH3) <400> 10 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 Xaa Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 11 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 11 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Arg Thr Lys Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 12 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (dpeg-dpeg-gammaGlu-CO(CH2)14CH3) <400> 12 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 13 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)3-CO(CH2)16CH3) <400> 13 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 14 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys((gammaGlu)5-CO(CH2)14CH3) <400> 14 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 15 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 15 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Tyr Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 16 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (dpeg-dpeg-gammaGlu-CO(CH2)14CH3) <400> 16 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Gly Thr Tyr 35 <210> 17 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 17 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 18 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys(CO(CH2)6CO2H) <400> 18 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 19 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(gammaGlu-dpeg-dpeg-(gammaGlu)3-CO(CH2)14CH3) <400> 19 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 20 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (dpeg-dpeg-gammaGlu-CO(CH2)14CH3) <400> 20 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Ala Thr Tyr 35 <210> 21 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys((gammaGlu)4-CO(CH2)14CH3) <400> 21 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 22 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 22 Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Tyr Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 23 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 23 Ser Cys Ser Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 24 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 24 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Ile Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 25 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys(gammaGlu-dpeg-dpeg-(gammaGlu)-CO(CH2)14CH3) <400> 25 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 26 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(gammaGlu-dpeg-dpeg-(gammaGlu)2-CO(CH2)14CH3) <400> 26 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 27 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)2-CO(CH2)14CH3) <400> 27 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 28 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)2-CO(CH2)14CH3) <400> 28 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Gln Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 29 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys((gammaGlu)3-CO(CH2)14CH3) <400> 29 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 30 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 30 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Glu Asn Phe Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 31 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys(CO(CH2)11CO2H) <400> 31 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 32 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 32 Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Glu Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 33 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 33 Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ser Asn Glu Leu 1 5 10 15 His Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 34 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 34 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ser Asn Glu Leu 1 5 10 15 His Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 35 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Leu <400> 35 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 36 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu-CO(CH2)14CH3) <400> 36 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 37 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 37 Ser Cys Asn Thr Ala Ser Cys Ala Thr Gln Arg Leu Ala Asn Tyr Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 38 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 38 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 39 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)2-Gly-CO(CH2)14CH3) <400> 39 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 40 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 40 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr His Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 41 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 41 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Arg Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 42 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys(gammaGlu-CO(CH2)14CH3) <400> 42 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 43 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 43 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 44 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 44 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 45 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 45 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 46 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Leu <400> 46 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa 1 5 10 15 Val Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 47 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu-CO(CH2)14CH3) <400> 47 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 48 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(dpeg-dpeg-gammaGlu-(CO(CH2)16CO2H)) <400> 48 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Gln Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 49 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 49 Ser Cys Ser Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Tyr Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 50 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Leu <400> 50 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 51 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 51 Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Glu Leu 1 5 10 15 Val Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 52 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 52 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ser Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 53 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)3-CO(CH2)14CH3) <400> 53 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 54 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 54 Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Glu Leu 1 5 10 15 His Arg Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 55 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(CO(CH2)16CO2H)) <400> 55 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 56 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)2-CO(CH2)16CH3) <400> 56 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 57 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 57 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 58 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 58 Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 59 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 59 Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Tyr Leu 1 5 10 15 Gln Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 60 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> D-Phe <400> 60 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 61 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 61 Ser Cys Asn Thr Ala Ser Cys Ala Thr Gln Arg Leu Ala Asn Tyr Leu 1 5 10 15 His Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 62 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 62 Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 63 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(gammaGlu-dpeg-dpeg-gammaGlu-CO(CH2)14CH3) <400> 63 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 64 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2(CO(CH2)18CO2H)) <400> 64 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 65 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2(CO(CH2)16CO2H)) <400> 65 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 66 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)4-CO(CH2)14CH3) <400> 66 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 67 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)3-CO(CH2)14CH3) <400> 67 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 68 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys((gammaGlu)2-CO(CH2)14CH3) <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys(2(gammaGlu)-CO(CH2)14CH3) <400> 68 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 69 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (dpeg-dpeg-gammaGlu-CO(CH2)14CH3) <400> 69 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Tyr Thr Tyr 35 <210> 70 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)2-CO(CH2)14CH3) <400> 70 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 71 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)14CH3) <400> 71 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 72 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys(CO(CH2)14CO2H)) <400> 72 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 73 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(gammaGlu-CO(CH2)18CO2H) <400> 73 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 74 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(CO(CH2)10CO2H)) <400> 74 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 75 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(CO(CH2)18CO2H) <400> 75 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 76 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(gammaGlu-CO(CH2)16CO2H) <400> 76 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 77 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)3-Gly-CO(CH2)14CH3) <400> 77 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 78 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (dpeg-dpeg-gammaGlu-CO(CH2)14CH3) <400> 78 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Pro Thr Tyr 35 <210> 79 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(CO(CH2)15CO2H) <400> 79 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 80 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(gammaGlu-CO(CH2)15CO2H) <400> 80 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 81 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(CO(CH2)12CO2H) <400> 81 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 82 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys((gammaGlu)2-CO(CH2)14CO2H) <400> 82 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 83 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys(CO(CH2)16CO2H) <400> 83 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 84 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(CO(CH2)8CO2H) <400> 84 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 85 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)18CO2H) <400> 85 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Tyr Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 86 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(CO(CH2)14CO2H) <400> 86 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 87 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys((gammaGlu)2-CO(CH2)16CO2H) <400> 87 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 88 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(gammaGlu-CO(CH2)14CO2H) <400> 88 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 89 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys(CO(CH2)18CO2H) <400> 89 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 90 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(CO(CH2)6CO2H) <400> 90 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 91 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)5-CO(CH2)14CH3) <400> 91 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 92 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys((gammaGlu)2-CO(CH2)18CO2H) <400> 92 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 93 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(CO(CH2)16CO2H) <400> 93 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 94 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)18CO2H) <400> 94 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 95 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> D-Lys(CO(CH2)18CO2H) <400> 95 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 96 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> D-Lys((gammaGlu)2-dpeg-CO(CH2)18CO2H) <400> 96 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 97 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> D-Lys((gammaGlu)2-CO(CH2)18CO2H) <400> 97 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 98 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> Lys(dpeg-dpeg-gammaGlu-CO(CH2)16CO2H) <400> 98 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 99 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (17)..(17) <223> Lys(dpeg-dpeg-gammaGlu-CO(CH2)16CO2H) <400> 99 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 Xaa Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 100 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> D-Lys((gammaGlu)2-dpeg-gammaGlu-CO(CH2)18CO2H) <400> 100 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 101 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> D-Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)18CO2H) <400> 101 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 102 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys(dpeg-dpeg-gammaGlu-CO(CH2)16CO2H) <400> 102 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 103 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)18CO2H) <400> 103 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 104 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)3-CO(CH2)16CO2H) <400> 104 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 105 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(dpeg-gammaGlu-CO(CH2)16CO2H) <400> 105 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Gln Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 106 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(gammaGlu-CO(CH2)16CO2H) <400> 106 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Gln Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 107 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (13)..(13) <223> Lys(gammaGlu-CO(CH2)14CH3) <400> 107 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Xaa Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 108 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (8)..(8) <223> Lys(gammaGlu-CO(CH2)14CH3) <400> 108 Ser Cys Asn Thr Ser Thr Cys Xaa Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 109 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(gammaGlu-CO(CH2)16CO2H) <400> 109 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 110 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(gammaGlu-CO(CH2)16CO2H) <400> 110 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Tyr Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 111 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 111 Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Tyr Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 112 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)18CO2H) <400> 112 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 113 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(dpeg-dpeg-gammaGlu-CO(CH2)16CO2H) <400> 113 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 114 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2-CO(CH2)18CO2H) <400> 114 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 115 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(CO(CH2)18CO2H) <400> 115 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 116 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(gammaGlu-CO(CH2)18CO2H) <400> 116 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 117 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2-CO(CH2)10CO2H) <400> 117 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 118 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2-CO(CH2)6CO2H) <400> 118 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 119 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(gammaGlu)-CO(CH2)18CO2H) <400> 119 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 120 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(CO(CH2)16CO2H) <400> 120 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 121 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (12)..(12) <223> Lys(gammaGlu)-CO(CH2)14CH3) <400> 121 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Xaa Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 122 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)14CH3) <400> 122 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 123 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys((gammaGlu)2-CO(CH2)14CH3) <400> 123 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 124 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)2-CO(CH2)20CH3) <400> 124 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 125 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> Lys(gammaGlu-CO(CH2)14CH3) <400> 125 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 126 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(dpeg-dpeg-gammaGlu-CO(CH2)16CO2H) <400> 126 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 127 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys <400> 127 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 128 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 128 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 129 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys <400> 129 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 130 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 130 Lys Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 131 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 131 Lys Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 132 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (15)..(15) <223> <400> 132 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 133 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (19)..(19) <223> <400> 133 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Val Lys Xaa Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 134 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> <400> 134 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 135 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (17)..(17) <223> <400> 135 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Xaa Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 136 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> N-MeLeu <400> 136 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ser Asn Phe Xaa 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 137 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> N-MeLeu <400> 137 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa 1 5 10 15 Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 138 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (14)..(14) <223> D-Asn <220> <221> MOD_RES <222> (16)..(16) <223> Lys((gammaGlu)5-(CO(CH2)14CH3) <400> 138 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Xaa Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Gly Thr Pro 35 <210> 139 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((gammaGlu)2-(CO(CH2)18CO2H) <400> 139 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Pro 35 <210> 140 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2-(CO(CH2)16CO2H) <220> <221> MOD_RES <222> (14)..(14) <223> D-Asn <400> 140 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Xaa Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Gly Thr Pro 35 <210> 141 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2-(CO(CH2)16CO2H) <220> <221> MOD_RES <222> (14)..(14) <223> D-Asp <400> 141 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Xaa Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Gly Thr Pro 35 <210> 142 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2-(CO(CH2)16CO2H) <220> <221> MOD_RES <222> (14)..(14) <223> D-Asp <400> 142 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Xaa Glu Leu 1 5 10 15 Arg His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Gly Thr Pro 35 <210> 143 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys((gammaGlu)2-(CO(CH2)14CO2H) <400> 143 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 144 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (acetyl) <400> 144 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 145 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (allyloxycarbonyl) <400> 145 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 146 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys((dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl) <400> 146 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 147 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu) <400> 147 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 148 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu-acetyl) <400> 148 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 149 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu-trityl) <400> 149 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 150 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu-tert-butyl) <400> 150 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 151 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu-gammaGlu) <400> 151 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 152 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu-gammaGlu-acetyl) <400> 152 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 153 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu-gammaGlu-trityl) <400> 153 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 154 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (16)..(16) <223> D-Lys (gammaGlu-gammaGlu-tert-butyl) <400> 154 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 155 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 155 Lys Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 156 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys(Fmoc) <400> 156 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 157 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu) <400> 157 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 158 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu-acetyl) <400> 158 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 159 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu-trityl) <400> 159 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 160 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu-tert-butyl) <400> 160 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 161 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu-gammaGlu) <400> 161 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 162 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu-gammaGlu-acetyl) <400> 162 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 163 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu-gammaGlu-trityl) <400> 163 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 164 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu-gammaGlu-tert-butyl) <400> 164 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu 1 5 10 15 Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 165 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu) <400> 165 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 166 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> MOD_RES <222> (1)..(1) <223> Lys (gammaGlu-gammaGlu) <400> 166 Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 167 <400> 167 000 <210> 168 <400> 168 000 <210> 169 <400> 169 000 <210> 170 <400> 170 000 <210> 171 <400> 171 000 <210> 172 <400> 172 000 <210> 173 <400> 173 000 <210> 174 <400> 174 000 <210> 175 <400> 175 000 <210> 176 <400> 176 000 <210> 177 <400> 177 000 <210> 178 <400> 178 000 <210> 179 <400> 179 000 <210> 180 <400> 180 000 <210> 181 <400> 181 000 <210> 182 <400> 182 000 <210> 183 <400> 183 000 <210> 184 <400> 184 000 <210> 185 <400> 185 000 <210> 186 <400> 186 000 <210> 187 <400> 187 000 <210> 188 <400> 188 000 <210> 189 <400> 189 000 <210> 190 <400> 190 000 <210> 191 <400> 191 000 <210> 192 <400> 192 000 <210> 193 <400> 193 000 <210> 194 <400> 194 000 <210> 195 <400> 195 000 <210> 196 <400> 196 000 <210> 197 <400> 197 000 <210> 198 <400> 198 000 <210> 199 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="Lys" or "D-Lys" or "His" or "Ile" <220> <221> VARIANT <222> (3)..(3) <223> /replace="Ser" <220> <221> VARIANT <222> (5)..(5) <223> /replace="Ala" <220> <221> VARIANT <222> (6)..(6) <223> /replace="Ser" <220> <221> VARIANT <222> (8)..(8) <223> /replace="Lys" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (12)..(12) <223> /replace="Lys" <220> <221> VARIANT <222> (13)..(13) <223> /replace="Ser" or "Glu" or "Lys" <220> <221> VARIANT <222> (14)..(14) <223> /replace="D-Asn" or "D-Asp" or "Tyr" or "Gln" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" or "D-Phe" or "Tyr" or "Ile" or "D-Lys" or "Lys" or "alpha-aminoisobutyric acid (Aib)" <220> <221> VARIANT <222> (16)..(16) <223> /replace="D-Lys" or "Leu" or "Aib" or "N-methyl leucine (N-MeL)" or "D-Leu" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" or "Arg" or "D-Lys" or "Lys" or "Aib" <220> <221> VARIANT <222> (18)..(18) <223> /replace="His" or "Arg" <220> <221> VARIANT <222> (19)..(19) <223> /replace="Aib" <220> <221> VARIANT <222> (20)..(20) <223> /replace="Aib" <220> <221> VARIANT <222> (22)..(22) <223> /replace="Glu" <220> <221> VARIANT <222> (29)..(29) <223> /replace="Arg" or "Lys" <220> <221> VARIANT <222> (31)..(31) <223> /replace="D-Lys" or "Asn" or "His" <220> <221> VARIANT <222> (35)..(35) <223> /replace="D-Glu" or "Asn" or "Lys" or "Gly" or "Ala" or "Tyr" or "Pro" <220> <221> VARIANT <222> (37)..(37) <223> /replace="Pro" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 199 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 200 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="Lys" or "D-Lys" or "His" or "Ile" <220> <221> VARIANT <222> (3)..(3) <223> /replace="Ser" <220> <221> VARIANT <222> (5)..(5) <223> /replace="Ala" <220> <221> VARIANT <222> (6)..(6) <223> /replace="Ser" <220> <221> VARIANT <222> (8)..(8) <223> /replace="Lys" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (12)..(12) <223> /replace="Lys" <220> <221> VARIANT <222> (13)..(13) <223> /replace="Ser" or "Glu" or "Lys" <220> <221> VARIANT <222> (14)..(14) <223> /replace="D-Asn" or "D-Asp" or "Tyr" or "Gln" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" or "D-Phe" or "Tyr" or "Ile" or "D-Lys" or "Lys" or "alpha-aminoisobutyric acid (Aib)" <220> <221> VARIANT <222> (16)..(16) <223> /replace="D-Lys" or "Leu" or "Aib" or "N-methyl leucine (N-MeL)" or "D-Leu" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" or "Arg" or "D-Lys" or "Lys" or "Aib" <220> <221> VARIANT <222> (18)..(18) <223> /replace="His" or "Arg" <220> <221> VARIANT <222> (19)..(19) <223> /replace="Aib" <220> <221> VARIANT <222> (20)..(20) <223> /replace="Aib" <220> <221> VARIANT <222> (22)..(22) <223> /replace="Glu" <220> <221> VARIANT <222> (29)..(29) <223> /replace="Arg" or "Lys" <220> <221> VARIANT <222> (31)..(31) <223> /replace="D-Lys" or "Asn" <220> <221> VARIANT <222> (35)..(35) <223> /replace="D-Glu" or "Asn" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 200 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 201 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="Lys" or "D-Lys" or "His" or "Ile" <220> <221> VARIANT <222> (3)..(3) <223> /replace="Ser" <220> <221> VARIANT <222> (5)..(5) <223> /replace="Ala" <220> <221> VARIANT <222> (6)..(6) <223> /replace="Ser" <220> <221> VARIANT <222> (8)..(8) <223> /replace="Lys" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (12)..(12) <223> /replace="Lys" <220> <221> VARIANT <222> (13)..(13) <223> /replace="Ser" or "Glu" or "Lys" <220> <221> VARIANT <222> (14)..(14) <223> /replace="D-Asn" or "D-Asp" or "Tyr" or "Gln" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" or "D-Phe" or "Tyr" or "Ile" or "D-Lys" or "Lys" or "Aib" <220> <221> VARIANT <222> (16)..(16) <223> /replace="D-Lys" or "Leu" or "Aib" or "N-MeL" or "D-Leu" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" or "Arg" or "D-Lys" or "Lys" or "Aib" <220> <221> VARIANT <222> (18)..(18) <223> /replace="His" or "Arg" <220> <221> VARIANT <222> (19)..(19) <223> /replace="Aib" <220> <221> VARIANT <222> (20)..(20) <223> /replace="Aib" <220> <221> VARIANT <222> (22)..(22) <223> /replace="Glu" <220> <221> VARIANT <222> (29)..(29) <223> /replace="Arg" or "Lys" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 201 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 202 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="D-Lys" or "Lys" <220> <221> VARIANT <222> (5)..(5) <223> /replace="Ala" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="D-Lys" or "Leu" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> VARIANT <222> (18)..(18) <223> /replace="His" or "Arg" <220> <221> VARIANT <222> (31)..(31) <223> /replace="Asn" <220> <221> VARIANT <222> (35)..(35) <223> /replace="Asn" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 202 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 203 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="D-Lys" or "Lys" <220> <221> VARIANT <222> (5)..(5) <223> /replace="Ala" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="D-Lys" or "Leu" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> VARIANT <222> (18)..(18) <223> /replace="His" or "Arg" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 203 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 204 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> VARIANT <222> (35)..(35) <223> /replace="Asn" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 204 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 205 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 205 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 206 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> VARIANT <222> (35)..(35) <223> /replace="Asn" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 206 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 207 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 207 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 208 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="D-Lys" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> VARIANT <222> (35)..(35) <223> /replace="Asn" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 208 Lys Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 209 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="D-Lys" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 209 Lys Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 210 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="Lys" or "D-Lys" or "His" or "Ile" <220> <221> VARIANT <222> (3)..(3) <223> /replace="Ser" <220> <221> VARIANT <222> (5)..(5) <223> /replace="Ala" <220> <221> VARIANT <222> (6)..(6) <223> /replace="Ser" <220> <221> VARIANT <222> (8)..(8) <223> /replace="Lys" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (12)..(12) <223> /replace="Lys <220> <221> VARIANT <222> (13)..(13) <223> /replace="Ser" or "Glu" or "Lys" <220> <221> VARIANT <222> (14)..(14) <223> /replace="D-Asn" or "D-Asp" or "Tyr" or "Gln" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" or "D-Phe" or "Tyr" or "Ile" or "D-Lys" or "Lys" or "alpha-aminoisobutyric acid (Aib)" <220> <221> VARIANT <222> (16)..(16) <223> /replace="D-Lys" or "Leu" or "Aib" or "N-methyl leucine (N-MeL)" or "D-Leu" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" or "Arg" or "D-Lys" or "Lys" or "Aib" <220> <221> VARIANT <222> (18)..(18) <223> /replace="His" or "Arg" <220> <221> VARIANT <222> (19)..(19) <223> /replace="Aib" <220> <221> VARIANT <222> (20)..(20) <223> /replace="Aib" <220> <221> VARIANT <222> (22)..(22) <223> /replace="Glu" <220> <221> VARIANT <222> (29)..(29) <223> /replace="Arg" or "Lys" <220> <221> VARIANT <222> (31)..(31) <223> /replace="D-Lys" or "Asn" <220> <221> VARIANT <222> (35)..(35) <223> /replace="D-Glu" or "Asn" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 210 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 211 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (10)..(10) <223> /replace="Ser" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> VARIANT <222> (35)..(35) <223> /replace="Asn" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 211 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 212 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <220> <221> VARIANT <222> (15)..(15) <223> /replace="Phe" <220> <221> VARIANT <222> (16)..(16) <223> /replace="Lys" or "D-Lys" <220> <221> VARIANT <222> (17)..(17) <223> /replace="Val" or "Gln" <220> <221> VARIANT <222> (35)..(35) <223> /replace="Asn" <220> <221> SITE <222> (1)..(37) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 212 Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu 1 5 10 15 His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val 20 25 30 Gly Ser Glu Thr Tyr 35 <210> 213 <400> 213 000 <210> 214 <400> 214 000 <210> 215 <400> 215 000 <210> 216 <400> 216 000 <210> 217 <400> 217 000 <210> 218 <400> 218 000 <210> 219 <400> 219 000 <210> 220 <400> 220 000 <210> 221 <400> 221 000 <210> 222 <400> 222 000 <210> 223 <400> 223 000 <210> 224 <400> 224 000 <210> 225 <400> 225 000 <210> 226 <400> 226 000 <210> 227 <400> 227 000 <210> 228 <400> 228 000 <210> 229 <400> 229 000 <210> 230 <400> 230 000 <210> 231 <400> 231 000 <210> 232 <400> 232 000 <210> 233 <400> 233 000 <210> 234 <400> 234 000 <210> 235 <400> 235 000 <210> 236 <400> 236 000 <210> 237 <400> 237 000 <210> 238 <400> 238 000 <210> 239 <400> 239 000 <210> 240 <400> 240 000 <210> 241 <400> 241 000 <210> 242 <400> 242 000 <210> 243 <400> 243 000 <210> 244 <400> 244 000 <210> 245 <400> 245 000 <210> 246 <400> 246 000 <210> 247 <400> 247 000 <210> 248 <400> 248 000 <210> 249 <400> 249 000 <210> 250 <400> 250 000 <210> 251 <400> 251 000 <210> 252 <400> 252 000 <210> 253 <400> 253 000 <210> 254 <400> 254 000 <210> 255 <400> 255 000 <210> 256 <400> 256 000 <210> 257 <400> 257 000 <210> 258 <400> 258 000 <210> 259 <400> 259 000 <210> 260 <400> 260 000 <210> 261 <400> 261 000 <210> 262 <400> 262 000 <210> 263 <400> 263 000 <210> 264 <400> 264 000 <210> 265 <400> 265 000 <210> 266 <400> 266 000 <210> 267 <400> 267 000 <210> 268 <400> 268 000 <210> 269 <400> 269 000 <210> 270 <400> 270 000 <210> 271 <400> 271 000 <210> 272 <400> 272 000 <210> 273 <400> 273 000 <210> 274 <400> 274 000 <210> 275 <400> 275 000 <210> 276 <400> 276 000 <210> 277 <400> 277 000 <210> 278 <400> 278 000 <210> 279 <400> 279 000 <210> 280 <400> 280 000 <210> 281 <400> 281 000 <210> 282 <400> 282 000 <210> 283 <400> 283 000 <210> 284 <400> 284 000 <210> 285 <400> 285 000 <210> 286 <400> 286 000 <210> 287 <400> 287 000 <210> 288 <400> 288 000 <210> 289 <400> 289 000 <210> 290 <400> 290 000 <210> 291 <400> 291 000 <210> 292 <400> 292 000 <210> 293 <400> 293 000 <210> 294 <400> 294 000 <210> 295 <400> 295 000 <210> 296 <400> 296 000 <210> 297 <400> 297 000 <210> 298 <400> 298 000 <210> 299 <400> 299 000 <210> 300 <211> 37 <212> PRT <213> Homo sapiens <400> 300 Lys Cys Asn Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Ala Ile Leu Ser Ser Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 301 <211> 37 <212> PRT <213> Rattus sp. <400> 301 Lys Cys Asn Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Val Arg Ser Ser Asn Asn Leu Gly Pro Val Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 302 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 302 Lys Cys Asn Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu 1 5 10 15 Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val 20 25 30 Gly Ser Asn Thr Tyr 35 <210> 303 <211> 32 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic polypeptide" <400> 303 Lys Cys Asn Thr Ala Thr Cys Val Leu Gly Arg Leu Ser Gln Glu Leu 1 5 10 15 His Arg Leu Gln Thr Tyr Pro Arg Thr Asn Thr Gly Ser Asn Thr Tyr 20 25 30 <210> 304 <211> 32 <212> PRT <213> Homo sapiens <400> 304 Cys Gly Asn Leu Ser Thr Cys Met Leu Gly Thr Tyr Thr Gln Asp Phe 1 5 10 15 Asn Lys Phe His Thr Phe Pro Gln Thr Ala Ile Gly Val Gly Ala Pro 20 25 30 <210> 305 <211> 32 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: salmon calcitonin" <400> 305 Cys Ser Asn Leu Ser Thr Cys Val Leu Gly Lys Leu Ser Gln Glu Leu 1 5 10 15 His Lys Leu Gln Thr Tyr Pro Arg Thr Asn Thr Gly Ser Gly Thr Pro 20 25 30 <210> 306 <211> 37 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: beta-calcitonin-gene-related peptide" <400> 306 Ala Cys Asn Thr Ala Thr Cys Val Thr His Arg Leu Ala Gly Leu Leu 1 5 10 15 Ser Arg Ser Gly Gly Met Val Lys Ser Asn Phe Val Pro Thr Asn Val 20 25 30 Gly Ser Lys Ala Phe 35 <210> 307 <211> 39 <212> PRT <213> Unknown <220> <221> source <223> /note="Description of Unknown: exenatide sequence" <400> 307 His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gin Met Glu Glu 1 5 10 15 Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro Ser 35 <210> 308 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="D-Lys" or "Lys" <220> <221> VARIANT <222> (5)..(5) <223> /replace="Ala" <220> <221> SITE <222> (1)..(7) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 308 Ser Cys Asn Thr Ser Thr Cys 1 5 <210> 309 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> VARIANT <222> (1)..(1) <223> /replace="Lys" or "D-Lys" <220> <221> SITE <222> (1)..(7) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 309 Ser Cys Asn Thr Ser Thr Cys 1 5 <210> 310 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <400> 310 Ser Cys Asn Thr Ser Thr Cys 1 5 <210> 311 <211> 10 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> MOD_RES <222> (1)..(10) <223> gammaGlu <220> <221> SITE <222> (1)..(10) <223> /note="This sequence may encompass 1-10 residues" <400> 311 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 <210> 312 <211> 11 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> MOD_RES <222> (1)..(10) <223> gammaGlu <220> <221> SITE <222> (1)..(10) <223> /note="This region may encompass 1-10 residues" <400> 312 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly 1 5 10 <210> 313 <211> 4 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> MOD_RES <222> (1)..(4) <223> gammaGlu <400> 313 Xaa Xaa Xaa Xaa One <210> 314 <211> 5 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> MOD_RES <222> (1)..(5) <223> gammaGlu <400> 314 Xaa Xaa Xaa Xaa Xaa 1 5 <210> 315 <211> 4 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> MOD_RES <222> (1)..(3) <223> gammaGlu <400> 315 Xaa Xaa Xaa Gly One <210> 316 <211> 11 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <220> <221> MOD_RES <222> (2)..(11) <223> gammaGlu <220> <221> SITE <222> (2)..(11) <223> /note="This region may encompass 1-10 residues" <400> 316 Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 <210> 317 <211> 4 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note="Description of Artificial Sequence: Synthetic peptide" <400> 317 Glu Glu Glu Glu One <210> 318 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> VARIANT <222> (1)..(1) <223> /replace="D-Lys" <220> <221> SITE <222> (1)..(7) <223> /note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 318 Lys Cys Asn Thr Ser Thr Cys 1 5

Claims (37)

An isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof:
X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TX ₃₇ -(OH/NH ₂ )(SEQ ID NO: 199), among the sequences,
X ₁ is S, K, k, H or I;
X ₃ is N or S;
X ₅ is S or A;
X ₆ is T or S;
X ₈ is A or K;
X ₁₀ is Q or S;
X ₁₂ is L or K;
X ₁₃ is A, S, E or K;
X ₁₄ is N, n, d, Y or Q;
X ₁₅ is E, F, f, Y, I, k, K or α-aminoisobutyric acid (Aib);
X ₁₆ is k, K, L, Aib, N -methyl leucine ( N -MeL) or 1;
X ₁₇ is H, V, Q, R, k, K or Aib;
X ₁₈ is K, H or R;
X ₁₉ is S or Aib;
X ₂₀ is S or Aib;
X ₂₂ is N or E;
X ₂₉ is P, R or K;
X ₃₁ is k, K, N or H;
X ₃₅ is e, E, N, K, G, A, Y or P; And
X ₃₇ is Y or P;
each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;
each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;
the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge;
provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K. The isolated polypeptide according to claim 1, comprising the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof:
X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 204), in the sequence,
X ₁ is S, K or k;
X ₁₀ is Q or S;
X ₁₅ is E or F;
X ₁₆ is L, K or k;
X ₁₇ is H, V or Q; And
X ₃₅ is E or N;
each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;
each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; And
The _{two cysteine residues of X 1} CNTSTC (SEQ ID NO: 309) are optionally further linked by a disulfide bridge. The isolated polypeptide according to claim 1, comprising the amino acid sequence of SEQ ID NO: 206 or a pharmaceutically acceptable salt thereof:
SCNTSTCATQRLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 206), in the sequence,
X ₁₅ is E or F;
X ₁₆ is L, K or k;
X ₁₇ is H, V or Q; And
X ₃₅ is E or N;
each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;
each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; And
The two cysteine residues of SCNTSTC (SEQ ID NO: 310) are optionally further linked by a disulfide bridge. According to claim 1,
SC*NTSTC*ATQRLANFkHKSSNNFGPILPPTKVGSETY-(NH2) (SEQ ID NO: 127);
SC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH2) (SEQ ID NO: 57);
SC*NTSTC*ATQRLANEKHKSSNNFGPILPPTKVGSETY-(NH2) (SEQ ID NO: 128);
SC*NTSTC*ATQRLANEkHKSSNNFGPILPPTKVGSETY-(NH2) (SEQ ID NO: 129); and
SC*NTSTC*ATQRLANFLVKSSNEFGPILPPTKVGSETY-(NH2) (SEQ ID NO:43)
An isolated polypeptide selected from the group consisting of. The amino acid sequence of claim 1 , wherein:
An isolated polypeptide comprising SC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH2) (SEQ ID NO: 57). The amino acid sequence of claim 1 , wherein:
An isolated polypeptide comprising SC*NTSTC*ATQRLANFkHKSSNNFGPILPPTKVGSETY-(NH2) (SEQ ID NO: 127). The isolated polypeptide of claim 1 , comprising the amino acid sequence: SC*NTSTC*ATQRLANEk*((γGlu) ₂ CO(CH ₂ ) ₁₄ CH ₃ )HKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 27). The isolated polypeptide according to claim 1, comprising the amino acid sequence of SEQ ID NO: 209 or a pharmaceutically acceptable salt thereof:
X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 209), in the sequence,
X ₁ is K or k;
X ₁₀ is Q or S;
X ₁₅ is E or F;
X ₁₆ is L, K or k; And
X ₁₇ is H, V or Q;
each K independently represents L -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer;
each k independently represents a D -lysine optionally covalently linked to a lipophilic substituent, optionally via a spacer; And
The _{two cysteine residues of X 1} CNTSTC (SEQ ID NO: 318) are optionally further linked by a disulfide bridge. According to claim 1,
An isolated polypeptide comprising the amino acid sequence of KC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH _{2 ) (SEQ ID NO: 130).} According to claim 1,
An isolated polypeptide comprising the amino acid sequence of K*((γGlu) ₂ (CO(CH ₂ ) ₁₈ CO ₂ H))C*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH _{2 )(SEQ ID NO: 64).} According to claim 1,
An isolated polypeptide comprising the amino acid sequence of K*((γGlu) ₂ (CO(CH ₂ ) ₁₆ CO ₂ H))C*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH _{2 ) (SEQ ID NO: 65).} According to claim 1,
An isolated polypeptide selected from the group consisting of KC*NTSTC*ATQRLANFLQKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 131). According to claim 1,
An isolated polypeptide selected from the group consisting of K*(γGluCO(CH ₂ ) ₁₆ CO ₂ H)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH _{2 (SEQ ID NO: 109).} The isolated polypeptide of claim 1 comprising an amino acid sequence selected from the group consisting of any one of SEQ ID NOs: 1-143. The isolated polypeptide of claim 1 , further comprising a lipophilic substituent and optionally comprising a spacer. The isolated polypeptide of claim 15 , further comprising a spacer and a lipophilic substituent of Formula VI:

during the meal,
Z is —CH ₃ or —CO ₂ H;
m is 4 to 24;
Y1 is selected from the group consisting of γGlu, Asp and Gly;
Y2 is selected from the group consisting of γGlu, Asp and Gly;
V is -[COCH ₂ (O(CH ₂ ) ₂ ) _t OCH ₂ NH]-, t is 1 to 8;
r is 1 to 8;
n1 is 0 to 10; And
n2 is 0 to 10. 16. The isolated polypeptide of claim 15, further comprising a spacer and a lipophilic substituent of Formula III:

during the meal,
Z is —CH ₃ or —CO ₂ H;
m is 4 to 24; And
n is 1 to 10. 17. The method of claim 16, wherein the lipophilic substituent -CO-(CH ₂ ) _m -Z is a spacer -(Y1) _n1 -(V) _r -(Y2) _n2 - of the lysine of the isolated polypeptide. An isolated polypeptide linked to the ε-amino group, wherein the spacer forms a bridge between the amino group of the disclosed polypeptide and the CO-group of the lipophilic substituent. 18. The isolated polypeptide of claim 17, wherein the lipophilic substituent -CO-(CH ₂ ) _m -Z is a spacer -(γGlu) _{n - (“(γGlu) n} ” disclosed as SEQ ID NO: 311). An isolated polypeptide linked to the ε-amino group of the lysine, wherein the spacer forms a bridge between the amino group of the disclosed polypeptide and the CO-group of the lipophilic substituent. 20. The isolated polypeptide of claim 18 or 19, wherein the lipophilic substituent is -CO-(CH ₂ ) _m -CO ₂ H. 20 . 21. The isolated polypeptide of claim 20, wherein m is 14 to 20. 20. The isolated polypeptide of claim 19, wherein the spacer is γGlu or 2(γGlu). 23. A pharmaceutical composition comprising the isolated polypeptide of any one of claims 1-22. 24. The pharmaceutical composition of claim 23, wherein the composition further comprises an insulinotropic polypeptide. 25. An osmotic delivery device comprising the isolated polypeptide of any one of claims 1-22 or the pharmaceutical composition of claims 23 or 24. 26. The method of claim 25,
an impermeable reservoir comprising inner and outer surfaces and first and second open ends.
a semipermeable membrane in sealing relationship with the first open end of the reservoir;
an osmotic engine in said reservoir and adjacent said semipermeable membrane;
a piston adjacent the osmosis engine, the piston forming a movable seal with an inner surface of the reservoir, the piston dividing the reservoir into a first chamber and a second chamber, the first chamber containing the osmosis engine , the piston,
a suspension formulation, wherein the second chamber contains the suspension formulation, the suspension formulation is flowable, the suspension formulation comprising the isolated polypeptide, and
a diffusion regulator inserted into the second open end of the reservoir, adjacent the suspension formulation;
comprising, an osmotic delivery device. 23. A method of treating obesity in a human subject, comprising providing weight loss in said human subject, or suppressing appetite in said human subject, comprising the isolated polypeptide of any one of claims 1-22. A method of treatment comprising administering to the subject the composition, the pharmaceutical composition of claim 23 or 24, or the osmotic device of claim 25 or 26. 25. A method of treating type 1 or type 2 diabetes in a human subject, comprising a pharmaceutical composition comprising the isolated polypeptide of any one of claims 1-22, a pharmaceutical composition of any one of claims 23 or 24, or A method of treatment comprising administering to the subject the osmotic device of claim 25 or 26 . 29. The method of claim 28, wherein the pharmaceutical composition comprising the isolated polypeptide is administered as an adjunct to administration of insulin. 30. The method of any one of claims 27-29, wherein the pharmaceutical composition comprising the isolated polypeptide is administered to the subject via implantation or injection. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 210:
X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 210 ), in the sequence,
X ₁ is S, K, k, H or I;
X ₃ is N or S;
X ₅ is S or A;
X ₆ is T or S;
X ₈ is A or K;
X ₁₀ is Q or S;
X ₁₂ is L or K;
X ₁₃ is A, S, E or K;
X ₁₄ is N, n, d, Y or Q;
X ₁₅ is E, F, f, Y, I, k, K or α-aminoisobutyric acid (Aib);
X ₁₆ is k, K, L, Aib, N -methyl leucine ( N -MeL) or 1;
X ₁₇ is H, V, Q, R, k, K or Aib;
X ₁₈ is K, H or R;
X ₁₉ is S or Aib;
X ₂₀ is S or Aib;
X ₂₂ is N or E;
X ₂₉ is P, R or K;
X ₃₁ is k, K or N; And
X ₃₅ is e, E or N;
each K independently represents L -lysine optionally covalently bound to a protecting group or a spacer optionally covalently bound to a protecting group;
each k independently represents a D -lysine optionally covalently bound to a protecting group or a spacer optionally covalently bound to a protecting group;
the _{two cysteine residues of X 1} CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge;
provided that _{if X 31} is N, then X ₃₅ is E, or if X ₃₅ is N, then X ₃₁ is K. 32. The method of claim 31,
SC*NTSTC*ATQRLANEkHKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 129);
SC*NTSTC*ATQRLANEk*(acetyl)HKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 144);
SC*NTSTC*ATQRLANEk*(allyloxycarbonyl)HKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 145); and
SC*NTSTC*ATQRLANEk*((dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl)HKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 146)
An isolated peptide selected from the group consisting of. 32. The method of claim 31,
SC*NTSTC*ATQRLANEk*HKSSNNFGPILPPTKVGSETY-NH2 (SEQ ID NO: 129);
SC*NTSTC*ATQRLANEk*(γGlu)HKSSNNFGPILPPTKVGSETY-NH2 (SEQ ID NO: 147);
SC*NTSTC*ATQRLANEk*(γGlu-acetyl)HKSSNNFGPILPPTKVGSETY-NH2 (SEQ ID NO: 148);
SC*NTSTC*ATQRLANEk*(γGlu-trityl)HKSSNNFGPILPPTKVGSETY-NH2 (SEQ ID NO: 149); and
SC*NTSTC*ATQRLANEk*(γGlu-tert-butyl)HKSSNNFGPILPPTKVGSETY-NH2 (SEQ ID NO: 150)
An isolated peptide selected from the group consisting of. 32. The method of claim 31,
SC*NTSTC*ATQRLANEk*(γGlu-γGlu)HKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 151);
SC*NTSTC*ATQRLANEk*(γGlu-γGlu-acetyl)HKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 152);
SC*NTSTC*ATQRLANEk*(γGlu-γGlu-trityl)HKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 153); and
SC*NTSTC*ATQRLANEk*(γGlu-γGlu-tert-butyl)HKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 154)
An isolated peptide selected from the group consisting of. 32. The method of claim 31,
KC*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 155); and
An isolated peptide selected from the group consisting of K*(Fmoc)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH _{2 (SEQ ID NO: 156).} 32. The method of claim 31,
K*(γGlu)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 157);
K*(γGlu-acetyl)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 158);
K*(γGlu-trityl)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 159); and
K*(γGlu-tert-butyl)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 160)
An isolated peptide selected from the group consisting of. 32. The method of claim 31,
K*(γGlu-γGlu)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 161);
K*(γGlu-γGlu-acetyl)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 162);
K*(γGlu-γGlu-trityl)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO:163); and
K*(γGlu-γGlu-tert-butyl)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 164)
An isolated peptide selected from the group consisting of.

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